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C-erbB-2 (human) clonal antibody (A24-V)

 
ALX-810-227-L001 1 ml 962.00 USD
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Product Specification

Alternative Name:ErbB2, HER-2, NEU proto-oncogene, CD340
 
Clone:A24-V
 
Host:Rabbit
 
Isotype:IgG
 
Immunogen:Synthetic peptide corresponding to the C-terminal region of human C-erbB-2.
 
UniProt ID:P04626
 
Species reactivity:Human
 
Applications:IHC (FS), IHC (PS)
 
Recommended Dilutions/Conditions:Immunohistochemistry (paraffin sections, 1:100-1:300)
Suggested dilutions/conditions may not be available for all applications.
Optimal conditions must be determined individually for each application.
 
Purity Detail:Major clone of IgGs obtained from immunoaffinity purified immunoglobulins corresponding to immunogenic peptide.
 
Formulation:Liquid. In 20mM TRIS/HCl, pH 8.0, containing 20mg/ml BSA and 0.05% sodium azide.
 
Use/Stability:Stable for at least 12 months after receipt when stored as recommended.
 
Handling:After opening, prepare aliquots and store at +4°C.
 
Shipping:Blue Ice Not Frozen
 
Long Term Storage:+4°C
 
Technical Info/Product Notes:Clonal Rabbit Antibody is a pure homogenous fraction of rabbit immunoglobulin (IgG) which corresponds to a unique linear epitope on the target protein. This epitope is designed after a detailed analysis of the antigen structure, regulatory post-translational modifications, protein/protein interaction domain and protein folding. The chosen sequence (peptide) is synthesized and the rabbit is immunized. The clone is then separated from the crude antiserum by "in vitro cloning technology". The resulting product is monospecific and characterized by superior specificity, affinity and avidity.
 
Protocol:IHC (paraffin sections) Protocol:
1. Deparaffinize the section in 3 changes of xylene, 5 minutes each.
2. Wash the section in 96%, 80% and 70% benzyl alcohol for 5 minutes each.
3. Rinse in distilled water.
4. Block the endogenous peroxidase by incubating the tissue in 3% hydrogen peroxide (H2O2) for 10 minutes.
5. Wash in distilled water.
6. For antigen retrieval use one of the following procedures:
A) Immerse the slide in TRIS-EDTA buffer, pH 9.0, 0.05% Tween- 20*, and incubate in microwave (600W) for 10 minutes, or
B) Immerse the slide in citrate buffer, pH 6.0, 0.05% Tween-20**, and incubate in microwave (600W) for 10 minutes.
7. Remove the staining to room temperature and let the slide to cool for 15 minutes.
8. Rinse in distilled water.
9. Wash in 0.05M TRIS-HCl , pH 7.6 buffer supplemented with 0.5% of Tween-20 (buffer A) for 5 minutes.
10. Incubate the section with primary antibody diluted in buffer A at the dilution 1:100- 1:300 for 1 hour in the closed wet chamber.
11. Wash twice 5 minutes with buffer A.
12. Apply the secondary antibody (the protocol depends on the supplier), and proceed to standard immunohistochemistry protocol (HRP - Peroxide - DAB).
13. Wash twice 5 minutes with buffer A.
14. Apply of chromogen (DAB), 10 minutes.
15. Wash in water - 10 minutes.
16. Stain in hematoxylin for 5 minutes.
17. Wash in water - 10 minutes.
18. Dehydrate the section in 2 changes of 96% benzyl alcohol for 5 minutes each.
19. Wash the section in 2 changes of xylene for 2 minutes each.
20. Mount the slide for observation.

* TRIS-EDTA Buffer (10mM TRIS Base, 1mM EDTA solution, 0.05% Tween-20, pH 9.0):
TRIS_____________1.21g;
EDTA_____________0.37g;
Distilled water______1000ml
Mix to dissolve in 700ml of distilled water. Adjust pH to 9.0 with 1M HCl and then add 0.5ml of Tween-20 and mix well. Adjust the final volume to 1 liter with distilled water.Store this solution at room temperature for 3 months or at +4°C for longer storage.

** Citrate Buffer (10mM Citric Acid, 0.05% Tween-20, pH 6.0):
Citric acid (anhydrous)______1.92g;
Distilled water____________1000ml;
Mix to dissolve in 700ml of distilled water. Adjust pH to 6.0 with 1M NaOH and then add 0.5ml of Tween-20 and mix well. Adjust the final volume to 1 liter with distilled water.Store this solution at room temperature for 3 months or at +4°C for longer storage.

 
810-227
Figure 1: Formalin-fixed and paraffin-embedded human breast ductal carcinoma tissue (4μm) stained with C-erbB-2 (human), cAb (A24-V) (Prod. No. ALX-810-227) shows strong membranous immunostaining of neoplastic cells.(Kindly performed and provided by Katarína Poliaková, MD and Lubomír Straka, MD, Ph. D. from Clinical Pathology Presov, Ltd., Presov, Slovakia)
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810-227

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