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CD45 (human) clonal antibody (E19-G) (Ready-to-Use)

 
ALX-810-410-L015 15 ml 410.00 USD
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Product Specification

Clone:E19-G
 
Host:Rabbit
 
Isotype:IgG
 
Immunogen:Synthetic peptide corresponding to a C-terminal sequence of human CD45.
 
UniProt ID:P08575
 
Species reactivity:Human
 
Applications:IHC (PS)
 
Purity Detail:Major clone of IgGs obtained from immunoaffinity purified immunoglobulins corresponding to immunogenic peptide.
 
Formulation:Liquid. Ready-to-use solution 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:Do not freeze.
 
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 for 5 minutes
6. For antigen retrieval: immerse the slide in the 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 (in citrate buffer, pH 6.0) for 15 minutes
8. Rinse in distilled water
9. Wash in 0.05M TRIS/HCl, pH 7.6, supplemented with 0.2% of Tween-20 (buffer A) for 5 minutes
10. Incubate the section with primary antibody (ready to use) 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. Rinse 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

*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.
 
810-410
Figure: Formalin-fixed and paraffin-embedded human tonsil tissue (4μm) stained with cAb to CD45 (human) (E19-G) shows strong positive immunostaining.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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810-410

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