Bcl-2 (human), cAb (P21-A)



ALX-810-224-R100	100 µl	165.00 USD

ALX-810-224-R500	500 µl	327.00 USD

ALX-810-224-L001	1 ml	544.00 USD

Product Specification

Concentration:	3mg/ml

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.

Clone:	P21-A

Isotype:	Rabbit IgG

Immunogen:	Synthetic peptide corresponding to an N-terminal sequence of human Bcl-2.

Specificity:	Recognizes human Bcl-2.

Application:	Immunohistochemistry (frozen sections, paraffin sections (1:100-1:200) Optimal conditions must be determined individually for each application.

Long Term Storage:	+4°C

Use/Stability:	Stable for at least 18 months after receipt when stored as recommended.

Handling:	After opening, prepare aliquots and store at +4°C.

Background / Technical Information:	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: immerse the slide in TRIS/EDTA buffer, pH 9.0, 0.05% Tween- 20, and incubate at 90 ˚C in water bath for 30 minutes. 7. Remove the staining to room temperature and let the slide to cool (in TRIS/EDTA buffer, pH 9.0) for 15 minutes. 8. Rinse in distilled water. 9. Wash in 0.05M TRIS/HCl, pH 7.6, 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- 1:200 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 the chromogen (DAB), 10 minutes. 15. Wash in water for 10 minutes. 16. Stain in hematoxylin for 5 minutes. 17. Wash in water for 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 ---------------------------- 1.21 g EDTA --------------------------- 0.37 g Distilled water ------–---–-- 1000 ml Mix to dissolve in 700ml of distilled water. Adjust pH to 9.0 with 1M HCI 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.

Figure: Formalin-fixed and paraffin-embedded human tonsil tissue (4μm) stained with cAb to Bcl-2 (human) (P21-A) (Prod. No. ALX-810-224) shows strong membrane immunostaining of folicular 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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