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Biotin Anti-Human CD45RA Antibody[HI100]

Cat:E-AB-F1052B
Manual MSDS

Price: $ 125

Price: $ 31

Size:
100μg 25μg
Quantity:
  • Application: FCM
  • Isotype: Mouse IgG2b, κ
  • Host: Mouse
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For research use only. Order now, ship in 3 days

Product Details
Background CD45RA is a 205-220 kD single chain type I glycoprotein. It is an exon 4 splice variant of the tyrosine phosphatase CD45. The CD45RA isoform is expressed on resting/naïve T cells, medullary thymocytes, B cells and monocytes. CD45RA enhances both T cell receptor and B cell receptor signaling. CD45 non-covalently associates with lymphocyte phosphatase-associated phosphoprotein (LPAP) on T and B lymphocytes. CD45 has been reported to be associated with several other cell surface antigens including CD1, CD2, CD3, and CD4. CD45 has also been reported to bind galectin-1. CD45 isoform expression can change in response to cytokines.
Alternate Names Receptor-type tyrosine-protein phosphatase C;Ptprc;L-CA;Ly-5;T200;CD45;
Swissprot
Clone No
Application
FCM
Host Mouse
Reactivity Human
Isotype Mouse IgG2b, κ
Isotype Control
Form Liquid
Conjugation
Biotin
Storage Buffer Phosphate buffered solution, pH 7.2, containing 0.09% stabilizer and 1% protein protectant.
Storage This product can be stored at 2-8°C for 12 months. Do not freeze.
Expiration date 12 months
Shipping Ice bag
  • Q1:What is the difference between Human CD45RA and CD45RO?

    CD45 is a receptor-type tyrosine phosphatase, also known as cluster of differentiation 45, that affects multiple signaling pathways, including T cell receptor (TCR) and B cell receptor (BCR) by regulating tyrosine phosphorylation levels. CD45 widely exists on the surface of hematopoietic cell membrane and is expressed on all leukocyte. Six isomers have been identified according to their extracellular epitopes, while CD45RA, CD45RB and CD45RO, have been identified on the surface of human cells. Then, two new subgroups of T cells can be identified by using this isomer molecule. CD45RA+ T cells are regarded as naive T cells (Tn) which means they are not stimulated by antigen , while CD45RO+ cells are known as memory T cells (Tm) been stimulated and differentiated by antigen.

  • Q2:Is it necessary to take blocking? Or for some specific samples?

    Blocking process is required when detecting macrophages, dendritic cells, NK cells. Fc receptors can be expressed on macrophages, dendritic cells, NK cells, etc. In the process of antibody staining in flow assay, Fc segment of FCM antibody will bind to Fc receptors on cell surface, end up with non-specific staining and lead to false positives signals. Antibodies can be incubated directly after blocking without washing.

  • Q3:Why centrifuge before use?

    During the transportation of antibodies, antibodies will stick to the tube wall or cap due to turbulence. So after receiving the antibodies, moderate centrifugation will collect the antibodies on the tube wall or cap to the bottom of the tube to avoid the loss of antibodies.

  • Q4:What auxiliary reagents are needed for staining?

    For samples with erythrocyte, ACK lysis buffer (E-CK-A105) is needed; For cells that are rich in Fc receptors, such as macrophages, Fc receptors blocking is necessary before staining with flow Antibody to reduce the non-specific signal. At present, we can provide human and mouse blocking agent, E-AB-F1236A Purified Anti-Human CD16 Antibody[3G8]. E-AB-F0997A Purified Anti-Mouse CD16/32 Antibody[2.4G2]. Cell staining buffer (E-CK-A107) is required in the process of cell staining. For the detection of intracellular indexes, a fixation & permeabilization kit is needed (E-CK-A109) ; and for the detection of intranuclear indexes, a specific staining kit (E-CK-A108) is required. Dead cell dyes are also used for flow cytometry of tissue samples such as tumors.

  • Q5:How should experimental groups/controls be set?

    Blank control: used to set the voltage of each channel. Isotype control: Isotype control antibodies are used as the basis for determining negative and positive cells. It is necessart as a gating helper especially for the indicators with low expression or continuous expression. Single color control: In a multicolor experiment, single color control is needed to adjust the fluorescence compensation if there is interference between the different channels. FMO control: FMO control, also known as fluorescence reduction control, refers to the multi-color experiment. FMO control is applied to observe the comprehensive effect of all related fluoresents to the target channel by removing the correspinding signal.

  • Q6:If there is only 1×10^5 cells instead of 1×10^6, can the antibody dosage be reduced?

    The amount of antibody is related to the incubation system, if the cell suspension volume is still 100 μL, the amount of antibody remains the same. Researchers can reduce the amount to save antibody in low cell number condition by reducing the cell suspension volume. It is recommended to use the recommended number of cells for the experiment, if the number of cells is too large, it will lead to insufficient antibody dosage, resulting in false negative; if the number of cells is too small, especially when detecting intracellular or intranuclear indicators, a large number of centrifugation operations will lose a lot of cells, resulting in insufficient number of cells for final detection.

  • Q7:How to dilute the Test package antibodies?

    The usage of test-package antibodies is well designed and verified, there is no need for an extra dilution. The usage amout of 1 test is 5 μL of antibodies per 100μL cell suspension (containing 1x10^6 cells).

  • Q8:What is the difference between the test-package and the weight-package of flow antibody products?

    The usage of test-package antibodies is well designed and verified there is no need for an extra dilution before use. The weight-package antibody has a higher concentration, and requires a titration process for a suitable usage amount.

  • Q9:Is it avaliable to store FCM antibodies at -20°C and thaw before use?

    The stability of the flow antibody can be different from each other based on differnet fluorescein. It is recommended to verify the freeze-thawed antibody through pre-experiments before determining. However, it is better to avoid the freeze-thawed cycling in any FCM antibody.

  • Q10:What does Isotype Control do? How to choose a suitable isotype control antibody?

    Isotype control antibodies are used as the basis for determining negative and positive cells. It is necessart as a gating helper especially for the indicators with low expression or continuous expression. The Isotype control was purified from the serum of non-immunized animals, it should be the same species source, same immunoglobulin and subtype, same fluorescein label, same dose and concentration as the stained monoclonal antibody.