Introduction
2-step plus is a two-step
immunohistochemical broad spectrum detection reagent. It polymerizes monovalent
Fab fragments of secondary antibody and enzyme, which
replaces the secondary antibody and tertiary antibody in traditional method,
can directly amplify the binding signal of antibody-antigen. This method not only retains the specific
binding ability of antibody with antigen, but also can effectively avoid space steric
hindrance caused by excessive polymer molecules. Compared with the traditional SP
three-step method, this kit has the characteristics of simple, rapid, high-sensitivity. This
system abandons the using of biotin, so it can avoid background staining by
endogenous biotin. It can be used in IHC, in which the primary antibody is monoclonal/polyclonal
antibody derived from goat. The unique polymer auxiliary agent for
macromolecular detection system can detect the binding primary antibody better.
Components
|
Cat
|
product
|
3 mL
|
6 mL
|
18 mL
|
Storage
|
|
E-IR-R212A
|
5% BSA
|
3 mL
|
6 mL
|
18 mL
|
2~8°C
|
|
E-IR-R212B
|
Polyperoxidase-anti-Goat IgG (Ready-to-Use)
|
3 mL
|
6 mL
|
18 mL
|
2~8°C
|
|
E-IR-R212C
|
3% H2O2
|
3 mL
|
6 mL
|
18 mL
|
2~8°C
|
|
Manual
|
|
One Copy
|
Sample dyeing
1. Dewax
and hydrate the paraffin section.
2. Make
thermal repair or digestion treatment to antigen of the tissue section if
necessary according to antigen/antibody situation.
3. Incubate with
E-IR-R212C (3% H2O2) for 10 min to eliminate endogenous peroxidase
activity. Wash with PBS or TBS, 2 min× 3.
4. Add
E-IR-R212A (5% BSA), incubate at room temperature for 30 min. Shake off any
excess liquid.
5. Add
primary antibody (Goat-IgG) with proper dilution ratio, incubate at 20~37°C for
1~2h or at 4°C overnight (then rewarm at 37°C for 30 min). Wash with PBS or TBS,
2 min× 3. Dry the section with absorbent paper.
6. Add
E-IR-R212B( Polyperoxidase-anti-Goat IgG), incubate at room temperature or 37°C
for 20 min. Wash with PBS or TBS, 2 min×3.
7. Color development with DAB.
8. Wash with deionized water to make the section counterstained,
dewatered, transparent, then seal the section.
Storage
Store at 2~8°C, shading light. Avoid of freezing. Valid for 12 months. The reagents are valid within 6 months after opening.
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