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CD44std (human) monoclonal antibody (SFF-304)

 
ALX-801-089-C100 100 µg Purified 284.00 USD
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Product Specification

Clone:SFF-304
 
Host:Mouse
 
Isotype:IgG1
 
Immunogen:Recombinant human CD44.
 
UniProt ID:P16070
 
Species reactivity:Human
 
Specificity:Does not distinguish between CD44 splice variants.
 
Applications:Flow Cytometry, IHC (FS), IHC (PS), WB
 
Recommended Dilutions/Conditions:Flow Cytometry: Recommended concentration for the unlabelled antibody is 2µg/ml and a 1:20 dilution of the labeled antibodies, as determined on HACAT cells.
Immunohistochemistry (frozen sections, paraffin sections): Can be used to stain both indirectly or directly acetone-fixed cryostat sections or cell smears. Recommended is the alkaline phosphatase-anti-alkaline-phosphatase (APAAP)
or peroxidase anti-peroxidase (PAP) procedures or the three stage immunoperoxidase technique on acetone-fixed cryostat sections. It may be used at a concentration of 2-10µg/ml for cryostat sections. Suitable for paraffin-embedded tissue after microwave treatment.
Western Blot: Recommended concentration: 2-10µg/ml.
Suggested dilutions/conditions may not be available for all applications. Optimal conditions must be determined individually for each application.
 
Formulation:Liquid. In PBS, pH 7.2-7.4.
 
Handling:Avoid freeze/thaw cycles.
 
Shipping:Shipped on Blue Ice
 
Short Term Storage:+4°C
 
Long Term Storage:-20°C
 
Protocol:Study protocol for testing the immunoreactivity of anti-CD44 antibodies on paraffin sections using wet autoclave and microwave pretreatment for antigen retrieval.

Patients material: Routinely formalin fixed and paraffin embedded biopsy material of 3 squamous cell carcinomas of the oral cavity, 3 breast carcinomas and colonic adenocarcinomas retrieved from the files of the Institute of Pathology, University of Münster

Handling of the sections: 1-2 µm thick paraffin sections were cut and mounted on poly-L-lysine coated glass slides. In cases of breast cancer different "glue" techniques were tested because of the frequent loss of sections during pretreatment procedures: the best results were achieved using either concentrated poly-L-lysine coated slides (without dilution layered over the precleaned glass slides) or by using Super Frost or Super Frost Plus slides (Menzel, Germany)

Dewaxing of the sections (as for routine immunohistochemistry):
2x15 minutes in xylene, 3 minutes in ethanol abs, 3 minutes in 90% ethanol, 3 minutes in 70% ethanol, 5 minutes in distilled water

Antigen retrieval using the wet autoclave protocol (Bankfalvi et al. J. Pathol. 174, 223 (1994)).
Dewaxed sections were immersed in sodium citrate buffer (0.01 M Na-citrate monohydrate, pH 6.0) in plastic Coplin jars and incubated in a Goessner Laborautoklav GLA 40-2 for 5 minutes.
 
ALX-801-089 IHC-2
Cervical tissue assayed with CD44std (human) monoclonal antibody (SFF-304) (ALX-801-089). Developed with POLYVIEW® PLUS HRP (anti-mouse) reagent (ENZ-ACC104) and HIGHDEF® DAB Chromogen/Substrate Set (ENZ-ACC105).
ALX-801-089 IHC-1
Cervical tissue assayed with CD44std (human) monoclonal antibody (SFF-304) (ALX-801-089). Developed with POLYVIEW® PLUS HRP (anti-mouse) reagent (ENZ-ACC104) and HIGHDEF® DAB Chromogen/Substrate Set (ENZ-ACC105).
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ALX-801-089 IHC-2 ALX-801-089 IHC-1

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