The essence of a clone number is the identifier for the hybridoma cell line that secretes a specific monoclonal antibody, serving as the unique "ID card" for that monoclonal antibody. The target determines which protein the antibody recognizes, while the clone number determines which epitope on that protein the antibody binds to, its binding affinity, and its tolerance to sample processing conditions. A large body of laboratory data and the standardization guidelines of the International Society for Advancement of Cytometry (ISAC) have confirmed that different clones against the same target can exhibit performance differences of orders of magnitude under identical experimental systems.
In practice, researchers often need to work across various experimental scenarios, each with its own requirements for clone selection. A "one-size-fits-all" approach often leads to chaotic flow cytometry data down the line.
This article will break down the key considerations for clone selection across six core application scenarios, helping experimenters choose the right clone at every step and avoid common pitfalls in flow cytometry experiments.
Table of Contents
1. Scenario 1: Live cell surface antigen detection: Must select clones that recognize extracellular conformational epitopes
2. Scenario 2: Low-abundance marker detection: High-affinity clones are indispensable
3. Scenario 3: Intracellular/nuclear antigen detection after fixation and permeabilization: Beware of loss of conformational epitopes
4. Scenario 4: High-parameter multicolor panel design: Validate spatial compatibility between clones
5. Scenario 5: Functional assays: Clone choice directly determines stimulation/protein transport inhibition effects
6. Scenario 6: Specially treated samples: e.g., CD4 detection following PMA stimulation
7. Summary and Key Recommendations for Clone Selection
01 Scenario 1: Live cell surface antigen detection: Must select clones that recognize extracellular conformational epitopes
Live cell surface antigen detection is one of the most common applications in basic flow cytometry-based immunophenotyping. The core principle is that the antibody must bind to the target under conditions where the cell membrane remains intact and the protein maintains its native conformation. Therefore, the selected clone must be capable of recognizing an extracellular conformational epitope on the target antigen, namely a three-dimensional spatial structure located on the extracellular region of the protein.
For example, in T cell analysis, different clones of CD3 antibodies show significant variation. Clones such as OKT-3 recognize the ε chain of the T cell receptor complex, UCHT1 primarily targets the extracellular domain of CD3, while CD3-12 mainly targets the intracellular segment of CD3. Their performance in live cell surface staining differs accordingly. Therefore, in live cell surface antigen detection, choosing a clone that has been flow-validated and explicitly labeled as suitable for live cell surface staining is the first step toward a successful experiment. (For detailed clone selection guidelines, refer to the summary table at the end of this article.)
02 Scenario 2: Low-abundance marker detection: High-affinity clones are indispensable
In stem cell research, rare cell subset analysis, and the detection of certain immune cell functional markers, researchers often face the challenge of extremely low target antigen expression levels. In such scenarios, affinity becomes a core consideration in clone selection. High-affinity clones can stably bind and generate strong signals even under conditions of low cell surface antigen expression; in contrast, low-affinity clones may bind weakly and be washed off during the washing steps, leading to weak signals or even false-negative results.
For example, in the tumor microenvironment, CD8 antigen expression is often low. If the selected clone has insufficient affinity, it becomes difficult to accurately identify these low-abundance antigens. A highly sensitive clone such as OKT-8 is therefore needed to better detect such low-proportion CD8+ T cells.
It is worth noting that high affinity does not equal high specificity. Some high-affinity clones are more prone to cross-reactivity, potentially binding to non-target antigens and reducing the signal-to-noise ratio of the assay. Therefore, in low-abundance marker detection, attention must be paid not only to affinity metrics (e.g., staining index, SI) but also to the specificity performance of the clone as validated in the literature. Both factors are indispensable.
03 Scenario 3: Intracellular/nuclear antigen detection after fixation and permeabilization: Beware of loss of conformational epitopes
When detecting intracellular or nuclear antigens (e.g., cytokines, transcription factors such as Foxp3, T-bet, etc.), samples must undergo fixation and permeabilization to allow antibodies to enter the cell membrane and even the nucleus. These treatments alter the native conformation of the antigen protein and may even disrupt the integrity of linear epitopes. Therefore, not all clones suitable for surface staining can be used for intracellular detection tasks.
In intracellular antigen detection, clone selection must prioritize antibodies that have been validated for fixation and permeabilization staining. Foxp3, as a key transcription factor, requires fixation and nuclear membrane permeabilization before it can be recognized by antibodies. If permeabilization is suboptimal or if the selected clone completely loses binding activity under fixation conditions, the staining signal will significantly deteriorate, potentially making the target protein undetectable. Therefore, choosing clones specifically optimized for nuclear protein detection and validated for intracellular staining is critical.
For example, among the recommended clones for nuclear Foxp3 detection, FJK-16s is recommended for mouse Foxp3 (widely cited in top journals such as Nature and Immunity), PCH101 is recommended for human Foxp3, and 4B10 is recommended for T-bet, all validated in fixation and permeabilization systems. Similarly, for surface antibodies used in combination with these nuclear markers, fixation-resistant clones should also be selected. (For detailed clone selection guidelines, refer to the summary table at the end of this article.)
Furthermore, the choice of fixation and permeabilization reagents themselves may affect clone compatibility. Some clones that recognize conformational epitopes may completely lose their binding ability after formaldehyde fixation. This means that when selecting clones, it is also necessary to choose a fixation and permeabilization protocol that is compatible with the chosen clone.
04 Scenario 4: High-parameter multicolor panel design: Validate spatial compatibility between clones
In multicolor panels, multiple antibodies are incubated simultaneously in the same sample. Different clones may exhibit steric hindrance or epitope competition, leading to reduced or even lost signals for certain markers. Therefore, in multicolor immunophenotyping experiments, the mutual compatibility between clones becomes a critical variable affecting experimental outcomes.
For example, in fine T cell immunophenotyping, each antibody clone must undergo systematic cross-compatibility validation to ensure stable staining in both peripheral blood mononuclear cells (PBMCs) and whole blood samples. When designing multicolor panels, it is necessary to systematically evaluate the compatibility of each clone. On one hand, check whether two clones recognize adjacent epitopes on the same antigen, which could lead to immunological competition; on the other hand, use co-incubation experiments to rule out unexpected steric hindrance effects. For instance, studies have shown that when a CD8α antibody (e.g., clone 11/295/33) is co-incubated with a CD8β antibody (e.g., PPT23 or PG164A), the binding efficiency of the latter can significantly decrease or even result in complete signal loss.
Furthermore, in multicolor panel design, the impact of fluorophore conjugation must also be given due consideration. The same clone may exhibit differences in signal intensity and background levels when conjugated to different fluorophores. Prioritize clone-fluorophore pairs that have extensive citation data in multicolor panels, and use single-staining controls and FMO controls to ultimately confirm whether the resolution meets experimental requirements.
05 Scenario 5: Functional assays: Clone choice directly determines stimulation/protein transport inhibition effects
In experimental designs involving cell stimulation, protein transport inhibition, or functional analysis, the choice of clone not only affects "detection performance" but also directly impacts "functional outcomes." Different clones of antibodies targeting the same molecule can produce completely distinct functional activities due to differences in their binding sites on the target molecule. Some clones may activate downstream signaling pathways, some may antagonize ligand binding, and others may have no effect on receptor function whatsoever.
For example, in T cell activation assays, different clones of CD3 antibodies exhibit significant differences in activation modes. The HIT3a clone is suitable for stimulating T cells in soluble form, whereas the UCHT1 clone requires pre-coating to exert its activating effect – the two are not interchangeable. In Natural Killer (NK) cell expansion experiments, the CB16 clone of anti-CD16 antibody efficiently promotes NK cell activation and expansion, while the MEM-154 clone shows essentially no activity. In PD-1/PD-L1-related research, the RMP1-14 clone is capable of blocking the PD-1/PD-L1 pathway and activating immune cells, whereas the RMP1-30 clone completely lacks such blocking function.
As long as the experimental goal involves eliciting a functional effect from the antibody – whether activation, blockade, or induction of internalization – one should not simply rely on clones intended for routine flow cytometry detection. Instead, detailed functional information about the selected clone must be verified in advance. Consulting literature reports on functional antibodies targeting the molecule of interest remains the most reliable approach.
06 Scenario 6: Specially treated samples: e.g., CD4 detection following PMA stimulation
Certain specialized treatment conditions can alter antigen expression characteristics, and in such cases, the choice of clone often becomes the critical breakthrough point for experimental success.
For example, in common Th1/Th2/Th17 functional studies, PMA (phorbol ester) stimulation induces partial internalization of surface CD4 on human T cells and accelerates its degradation in lysosomes. This leads to a significant decrease in positive signals when using conventional CD4 antibodies for detection, making it impossible to distinguish between positive and negative cell populations. In such cases, it is necessary to select a clone such as SK3, which is less affected by internalization, to still clearly distinguish CD4-positive from CD4-negative cell populations after stimulation.
In summary, when sample processing causes changes in the expression characteristics of the target molecule, the suitability of conventional clones may "fail," and alternative clones that bypass the internalization epitope need to be identified.
07 Summary and Key Recommendations for Clone Selection
In summary, when selecting antibodies for flow cytometry experiments, one should adhere to the principle of prioritizing application-specific validation, obtain recommendations from authoritative sources, and perform retrospective verification using literature databases. Be vigilant about various experimental pitfalls related to clone selection, and make clone choice a primary consideration in experimental design to ensure flow cytometry data quality and the verifiability and reproducibility of scientific research results.
We have summarized the commonly used antibody clone selections for T cell immunophenotyping in both humans and mice for your reference, as shown in the table below.
Table 1. Clone Selection for Human T Cells Immunophenotyping Antibodies
|
Target |
Clone No. |
Feature |
Application |
|
CD3 |
OKT3 |
CD3ε extracellular domain epitope, overlapping and competing with UCHT1/SK7 epitopes; extremely strong activation capability, triggers T cell proliferation and substantial secretion of IL-2/IFN-γ upon crosslinking; suitable for live cell detection, readily inactivates after fixation |
Gold standard for T cell functional assays; T cell in vitro activation/proliferation experiments; CAR-T production; functional stimulation; transplantation immunology research. |
|
UCHT1 |
CD3ε extracellular domain epitope, conformation-dependent, stains both surface and intracellular CD3; weak activation capability, does not induce strong proliferation/cytokine storm; suitable for live cells, fixation/permeabilization (mild conditions), and frozen sections. |
First choice for routine flow cytometry + intracellular staining; immunophenotyping; intracellular staining; first choice for multicolor panels. |
|
|
SK7 |
CD3ε extracellular domain epitope, conformationally stable, good fixation tolerance; moderate activation capability, soluble antibody requires monocyte assistance to exhibit mitogenic activity; suitable for live cells and fixed cells, formalin-fixed paraffin-embedded (FFPE) sections, and frozen sections. |
Clinical grade, first choice for standardized detection + paraffin IHC; immunohistochemistry (FFPE/frozen); T cell sorting; stable multicolor compensation. |
|
|
HIT3a |
CD3ε extracellular domain epitope, recognizes only native conformation of surface CD3, does not bind intracellular CD3; weak activation capability, suitable for activation-free phenotyping; suitable for live cells, ineffective after permeabilization. |
Surface-specific, avoids intracellular interference, avoids intracellular signaling interference, high-purity T cell sorting. |
|
|
MEM-57 |
Conformational epitope of CD3δ-ε/γ-ε complexes, does not recognize free CD3ε; moderate activation capability, dependent on TCR cross-linking; suitable for live cells and mild fixation. |
TCR/CD3 complex integrity detection, immunoprecipitation, conformational studies. |
|
|
CD3-12 |
Linear epitope of the CD3ε intracellular domain, highly conserved, broad cross-species reactivity; no activation capability; suitable for strong fixation/permeabilization, paraffin sections, and Western blot. |
Exclusive for intracellular CD3 staining, thymocyte development analysis, cross-species immunohistochemistry, WB. |
|
|
CD4 |
SK3/Leu3a |
CD4 extracellular domain D1 region, conformation-dependent, partially overlapping with OKT4 epitope, partially blocks binding of the SK3 clone antibody; weak activation capability, does not induce strong proliferation or cytokine secretion in CD4+ T cells; resistant to formaldehyde fixation, suitable for live cells, mild fixation/permeabilization samples, formalin-fixed paraffin-embedded (FFPE) samples, and frozen sections. |
General first choice for both clinical and research use; insensitive to internalization (detectable even after PMA stimulation); cytokine detection in Th1/Th2/Th17 etc.; HIV infection research; immunophenotyping. |
|
OKT-4 |
CD4 extracellular domain D1 region, conformation-dependent, partially overlapping with RPA-T4 epitope, competitively binds to CD4 molecule; extremely strong activation capability, efficiently triggers CD4+ T cell proliferation and substantial secretion of cytokines such as IL-2 and IFN-γ upon crosslinking; readily inactivates after fixation, preferentially suitable for live cells, signal decreases significantly after mild fixation, completely inactivated after strong fixation (e.g., formalin), not suitable for fixed/permeabilized samples or paraffin section detection. |
Classic activating type, dedicated for functional assays; routine immunophenotyping; T cell functional studies; HIV research; CD4+ T cell proliferation, differentiation, and signaling studies. |
|
|
RPA-T4 |
CD4 extracellular domain D1 region, conformation-dependent, partially overlapping with OKT4 epitope, partially blocks binding of the SK3 clone antibody; moderate activation capability, mild activating effect when used alone, weakly induces CD4+ T cell proliferation upon crosslinking, possesses functional blocking properties, can block HIV binding to CD4; poor tolerance to formaldehyde fixation, suitable for live cells, mildly fixed samples, immunocytochemistry/immunofluorescence samples, applicable to multiple platforms including flow cytometry, immunoprecipitation, and Western blot. |
Multi-scenario compatible, functional blocking type: strong multi-platform adaptability, suitable for HIV infection mechanism research, CD4+ T cell functional regulation, and multi-platform combined detection assays; limited cross-species reactivity, primarily applicable to human and chimpanzee samples. |
|
|
13B8.2 |
Linear epitope of the CD4 intracellular domain, highly conserved, recognizes free CD4 intracellular fragments and intracellularly expressed CD4 molecules, broad cross-species reactivity (recognizes CD4 from human, monkey, dog, and other species); no activating effect, does not bind to cell surface CD4 due to recognition of intracellular epitope; fixation-resistant, suitable for intracellular CD4 detection scenarios, compatible with strong fixation/permeabilization samples, no signal in live cell detection. |
Dedicated for intracellular/cross-species use: good cross-species compatibility, stable recognition of intracellular epitope, suitable for thymocyte development analysis (tracking early CD4+ T cells), intracellular CD4 expression detection, cross-species immunological studies, and Western blot experiments, avoids interference from surface CD4 signal. |
|
|
CD8 |
SK1 |
Recognizes the extracellular conformational epitope of CD8α; the greatest advantage is that it does not compete with MHC tetramers for binding, provides stable signal intensity, has good tolerance for mild fixation with 1-4% PFA, is compatible with live cell and fixed sample staining, weak activating properties do not interfere with cellular functional assays, exhibits stable compensation and low non-specific staining in multicolor flow cytometry. |
Most universal clone, first choice for routine T cell phenotyping (CD3/CD4/CD8 combination staining), suitable for standardized detection in whole blood, PBMC, and other samples; can be used for fixed samples followed by intracellular staining experiments; also suitable for multicolor flow cytometry panel construction, widely applied in basic research and clinical routine detection. |
|
OKT-8 |
Recognizes the extracellular epitope of CD8α; exhibits very strong signal intensity and high sensitivity in live cell staining; can synergistically enhance signal in MHC tetramer staining for certain antigens; however, has poor fixation tolerance —: signal significantly decreases after fixation treatment, and population resolution may even become blurred. |
Suitable for high-sensitivity detection requirements, such as detection of cell subsets with low CD8 expression and rare CD8+ T cells; widely used in CAR-T and clinical quality control, serving as a reliable marker. |
|
|
Hit8a |
Recognizes a unique extracellular epitope of CD8α, completely non-competitive with MHC tetramer binding, making it the exclusive preferred clone for antigen-specific CD8+ T cell detection; good performance in live cell staining, moderate tolerance to mild fixation, but suboptimal performance in intracellular staining with moderate signal intensity. |
Tetramer-friendly clone, gold standard for co-staining of MHC tetramers with CD8, widely used in the detection of antigen-specific CD8+ T cells in tumors and infectious diseases; suitable for multicolor MHC tetramer panel construction, enabling simultaneous detection of multiple antigen-specific CD8+ T cell subsets; first choice for antigen-specific T cell functional assays in basic research. |
|
|
UCHT-4 |
Recognizes the extracellular conformational epitope of CD8α. This antibody is immunized with human thymocytes and peripheral blood T cells, recognizes both CD8αα homodimers and CD8αβ heterodimers, and exhibits good reactivity in both thymocytes and mature T cells; stable signal intensity in live cell staining with low non-specific staining; sensitive to epitopes in conventionally formalin-fixed samples with weak fixation tolerance —: partial signal is retained under mild fixation conditions; good stability in multicolor flow cytometry compensation, with signal intensity slightly weaker than SK1 and OKT-8. |
Suitable for basic research and some clinical routine CD8+ T cell phenotyping (whole blood, PBMC samples); can be used in combination with CD3 and CD4 antibodies for T cell subset analysis; compatible with multiple experimental methods including flow cytometry, immunofluorescence (IF), and immunohistochemistry (IHC), particularly suitable for experiments requiring both flow cytometry and immunofluorescence detection, enabling multi-use of a single antibody and reducing experimental variability. |
|
|
HI149 |
Recognizes a specific extracellular epitope of CD8α, exhibits high specificity in live cell staining with extremely low non-specific staining, no competitive binding with MHC tetramers, good compatibility with multicolor flow cytometry panels, moderate signal intensity; has some tolerance to mild fixation (1-4% PFA), can be used for staining of short-term fixed samples with good signal retention after fixation, excellent batch-to-batch stability, reacts only with human CD8 molecules with no cross-species reactivity. |
Primarily used for routine CD8+ T cell phenotyping and CD8+ T cell subset screening in basic research, suitable for flow cytometry detection of whole blood, PBMC, and sorted cells; suitable for multicolor flow cytometry panel construction, particularly compatible with high-signal antibodies to reduce compensation interference; can be used for positive selection of CD8+ T cells with good post-sort cell viability. |
|
|
B9.11 |
Recognizes a stable conformational epitope of CD8α; has good tolerance for formalin-fixed paraffin-embedded (FFPE) samples, excellent performance in live cell staining, retains complete signal after mild fixation, good compatibility with multicolor flow cytometry panels, signal intensity slightly weaker than SK1 and OKT-8, but strong batch-to-batch stability, suitable for clinical standardized detection. |
Primarily used for clinical routine CD8 phenotyping by flow cytometry, suitable for batch testing of clinical samples such as whole blood and PBMC; can also be used for immunohistochemistry (frozen sections, paraffin sections), enabling antibody consistency between flow cytometry and IHC experiments and reducing experimental variability; can also be used for routine phenotyping in basic research, suitable for experiments requiring high batch-to-batch stability. |
Table 2. Clone Selection for Mouse T/B/NK Immunophenotyping Antibodies
|
Target |
Clone No. |
Feature |
Application |
|
CD3 |
145-2C11 |
Recognizes the extracellular epitope of the mouse CD3ε chain. The epitope recognized by this clone depends on the conformational association of the CD3ε chain with the δ or γ chain; it cannot recognize cells expressing only the isolated ε chain, and may fail to detect some NK tumors. Extremely strong signal intensity and very high specificity with no cross-species reactivity (does not react with rat leukocytes); good tolerance to mild fixation with 1-4% PFA, compatible with live cell and fixed sample staining, stable compensation in multicolor flow cytometry; can initiate T cell activation signaling by binding to the CD3ε chain, suitable for both flow cytometry detection and functional assays; applicable to commonly used mouse strains such as C57BL/6 and BALB/c. |
Gold standard clone, first choice for mouse CD3 flow cytometry detection, suitable for routine T cell phenotyping (CD3/CD4/CD8 combination staining), compatible with various mouse sample types including whole blood, spleen, thymus, lymph nodes, PBMC, and tumor-infiltrating T cells; can be used for subsequent intracellular staining of fixed samples (e.g., detection of intracellular cytokines and transcription factors in T cells); also suitable for T cell sorting, activation, exhaustion, and blockade experiments, widely used in basic research (tumor immunology, autoimmune diseases, infection models) and some preclinical mouse studies; can also be used for multi-methodology validation including immunohistochemistry (frozen sections) and immunoprecipitation (IP). |
|
17A2 |
Recognizes a conserved conformational epitope on the extracellular domain of the mouse CD3 molecule, low non-specific staining, highly compatibility with multicolor flow cytometry panels. Has some tolerance to mild fixation, but signal slightly decreases after strong fixation (prolonged fixation with 4% PFA); stable signal in live cell staining; can initiate T cell activation and proliferation by binding to the CD3 molecule; applicable to commonly used mouse strains such as C57BL/6 and BALB/c. |
Routine mouse T cell phenotyping analysis, suitable for flow cytometry detection of mouse splenocytes, thymocytes, peripheral blood, and lymphoid tissue samples; a commonly used alternative clone for routine T cell phenotyping in basic research. Can be used for T cell subset identification, immune response studies, and tumor immune microenvironment analysis; suitable for multicolor flow cytometry panel construction, particularly compatible with combined detection of antibodies such as CD4, CD8, and PD-1; can also be used for immunofluorescence (IF) experiments. Not suitable for detection of rare T cell subsets with low CD3 expression. |
|
|
CD4 |
GK1.5 |
Recognizes an extracellular epitope of CD4, competes with RM4-5 for the same epitope, does not compete with RM4-4; high brightness, good specificity, low background; signal is retained after mild fixation with 1-2% PFA, suitable for combined surface and intracellular co-staining (Treg/Th17); signal decreases significantly after strong fixation/permeabilization; applicable to commonly used mouse strains such as C57BL/6 and BALB/c. |
The most commonly used clone for flow cytometry detection: routine CD4+ T cell phenotyping; Treg (CD25/Foxp3); Th1/Th2/Th17 multicolor panels; spleen, lymph nodes, whole blood, tumor-infiltrating lymphocytes; sorted cells can still be used for functional assays (proliferation/cytokines). |
|
RM4-5 |
Recognizes an extracellular epitope of CD4, competes with GK1.5, does not compete with RM4-4; higher affinity and better signal-to-noise ratio compared to GK1.5; exhibits good tolerance to mild fixation, with better signal retention in intracellular staining than GK1.5. |
First choice flow cytometry detection antibody for in vivo experiments: high brightness, suitable for multicolor panels and rare subsets (low CD4 expression, TILs, etc.). |
|
|
RM4-4 |
Recognizes a different extracellular epitope of CD4, with no competition against GK1.5 or RM4-5; moderate-to-high affinity with clean background; resistant to mild fixation (1%-4% PFA), can be used for surface staining followed by fixation and mild permeabilization, suitable for combined Treg/intracellular cytokine staining; fixation tolerance of RM4-4 is better than GK1.5 but slightly weaker than RM4-5. |
Enables simultaneous detection when paired with appropriate fluorophores, making it an ideal choice for T cell sorting; also used for dual-clone staining in the same tube (e.g., GK1.5 + RM4-4) to verify authentic CD4 expression and rule out epitope loss; thymocyte development (CD4/CD8 double-positive stage). |
|
|
YTS191 |
Recognizes a membrane-proximal extracellular epitope of CD4, blocks CD4–MHC II binding, competes with GK1.5; severe signal loss after fixation, suitable only for live cell staining. |
Classic depleting antibody, more suitable for functional assays (in vitro T cell activation inhibition, co-stimulation blockade experiments) rather than phenotyping; commonly used for in vivo CD4 blockade/depletion models (autoimmunity, transplant tolerance); studies on CD4 molecular conformation and functional relationships; not suitable for high-sensitivity phenotyping analysis. |
|
|
CD8 |
53-6.7 |
Recognizes an extracellular epitope of mouse CD8α, recognizes both CD8αα and CD8αβ, does not significantly compete with MHC-I tetramers, suitable for antigen-specific CD8+ T cell detection; no significant T cell activation, safe for phenotypic staining; good fixation tolerance, tolerates mild fixation with 1%-4% PFA, compatible with routine permeabilization and intracellular cytokine/transcription factor co-staining. |
Scientific research gold standard, all-purpose first choice: universal for all sample types including spleen/lymph nodes/thymus/TIL/peripheral blood; TIL, tumor-infiltrating CD8+ T cells, exhausted T cell detection; intracellular staining (IFN-γ, Granzyme B, Perforin); multicolor panels; cell sorting; long-term fixation before flow cytometry acquisition. |
|
YTS169.4/YTS-169 |
Recognizes a membrane-proximal functional epitope of CD8α, blocks CD8–MHC I binding; poor fixation tolerance, suitable only for fresh live cells, signal significantly decreases after fixation and permeabilization, cannot be used for intracellular staining. |
Low endotoxin level, specifically designed for in vivo experiments, high specificity; suitable for in vitro blockade, in vivo CD8+ T cell depletion/elimination models, and in vitro CD8 functional blockade assays. |
|
|
NLDC-145 |
Recognizes a specific epitope on the CD8β chain, only recognizes CD8αβ heterodimers, does not recognize CD8αα double-positive subsets in the intestine or thymus, nor NK-like CD8 cells; moderate fixation tolerance, compatible with mild PFA fixation, not suitable for strong permeabilization. |
Precisely distinguishes conventional peripheral mature CD8αβ+ cytotoxic T cells, will miss CD8αα cells, cannot be used as a universal total CD8 marker; can exclude unconventional CD8αα subsets for fine subset analysis; mucosal immunity, thymic development subclassification studies. |
