33-6700

Campos D, Freitas D, Gomes J, Magalhães A, Steentoft C, Gomes C, Vester-Christensen MB, Ferreira JA, Afonso LP, Santos LL, Pinto de Sousa J, Mandel U, Clausen H, Vakhrushev SY, Reis CA
Molecular & cellular proteomics : MCP 2015 Jun;14(6):1616-29

Co-expression of CD133, CD44v6 and human tissue factor is associated with metastasis and poor prognosis in pancreatic carcinoma.

Chen K, Li Z, Jiang P, Zhang X, Zhang Y, Jiang Y, He Y, Li X
Oncology reports 2014 Aug;32(2):755-63

Leuprorelin acetate affects adhesion molecule expression in human prostate cancer cells.

Angelucci C, Lama G, Iacopino F, Sica G
International journal of oncology 2011 Jun;38(6):1501-9

De novo expression of CD44 variants in sporadic and hereditary gastric cancer.

da Cunha CB, Oliveira C, Wen X, Gomes B, Sousa S, Suriano G, Grellier M, Huntsman DG, Carneiro F, Granja PL, Seruca R
Laboratory investigation; a journal of technical methods and pathology 2010 Nov;90(11):1604-14

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts of KARPAS 299 (Lane 1), HEL 92.1.7 (Lane 2), THP-1 (Lane 3), U-87 MG (Lane 4) and IMR-32 (Lane 5) and PC-3 (Lane 6). The blots were probed with Anti-CD44V Mouse Monoclonal Antibody (Product # 33-6700, 2 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conj µgate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). Bands of ~ 81 and 77 kDa band corresponding to CD44V was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of CD44v6 was performed using 90% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with CD44v6 (MA54) Mouse Monoclonal Antibody (Product # 33-6700) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conj µgate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing localization in the membrane. Panel e shows the no primary antibody control. The images were captured at 60X magnification.