CD44v6 Antibody Staining Protocol for Immunohistochemistry


Description: Mouse monoclonal antibody derived from a fusion of mouse myeloma cells (SP2/0) and spleen cells from a mouse immunized with a human chimeric fusion protein CD44v3-10-Fc. Purified from tissue culture supernate by affinity chromatography on a protein-A matrix.


Primary Antibody

Name: CD44v6 Antibody

Clone: 2F10, Mouse anti-Human

Supplier: R&D Systems

Catalog Number: BBA13

Dilution: 1:500 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60 min/room temperature

Antigen Retrieval
Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95-100 ºC/room temperature
Heat/Cool Time: 20 minutes/20 minutes

Detection Methods
Standard Method: ABC Method or LSAB Method
Enhanced Method: Polymeric Methods

Chromogen Substrate
Reagent: DAB
Incubation Time/Temperature: 1-3 minutes/room temperature

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Cell membrane
Images: Search image

Additional Information:
Tissue Type: Colon
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Human PBMCs, colon, breast carcinoma, pancreatic adenocarcinomas
Negative Control: Omit primary antibody, isotype control, absorption control
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary


Not need to perform antigen retrieval on acetone fixed frozen sections



1.   Morris SF, O'Hanlon DM, McLaughlin R, McHale T, Connolly GE, Given HF. (2001) The prognostic significance of CD44s and CD44v6 expression in stage two breast carcinoma: an immunohistochemical study. Eur J Surg Oncol. 27(6):527-31. PubMed Abstract

2.   Castella EM, Ariza A, Ojanguren I, Mate JL, Roca X, Fernandez-Vasalo A, Navas-Palacios JJ (1996) Differential expression of CD44v6 in adenocarcinoma of the pancreas: an immunohistochemical study. Virchows Arch. 429(4-5):191-5. PubMed Abstract

3.   Rosenberg WM, Prince C, Kaklamanis L, Fox SB, Jackson DG, Simmons DL, Chapman RW, Trowell JM, Jewell DP, Bell JI (1995) Increased expression of CD44v6 and CD44v3 in ulcerative colitis but not colonic Crohn's disease. Lancet. 345(8959):1205-9. PubMed Abstract