- 730046 - Invitrogen Antibodies CBX5 antibody

Submitted references Assessment of a method to characterize antibody selectivity and specificity for use in immunoprecipitation.

Marcon E, Jain H, Bhattacharya A, Guo H, Phanse S, Pu S, Byram G, Collins BC, Dowdell E, Fenner M, Guo X, Hutchinson A, Kennedy JJ, Krastins B, Larsen B, Lin ZY, Lopez MF, Loppnau P, Miersch S, Nguyen T, Olsen JB, Paduch M, Ravichandran M, Seitova A, Vadali G, Vogelsang MS, Whiteaker JR, Zhong G, Zhong N, Zhao L, Aebersold R, Arrowsmith CH, Emili A, Frappier L, Gingras AC, Gstaiger M, Paulovich AG, Koide S, Kossiakoff AA, Sidhu SS, Wodak SJ, Gräslund S, Greenblatt JF, Edwards AM
Nature methods 2015 Aug;12(8):725-31

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of CBX5 was achieved by transfecting HeLa cells with CBX5 specific validated siRNA (Silencer® select Product # s23885). Western blot analysis was performed using whole cell lysates from the CBX5 knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti- CBX5 Recombinant Mouse monoclonal Antibody (Product # 730046, 2 µg/mL) and Biotin conjugated streptavidin Poly HRP (Product # 21140, 1:10000 dilution). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to CBX5.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of NTERA-2 (Lane 1), HeLa (lane 2), HEK-293 (Lane 3), K-562 (Lane 4) and A549 (Lane 5). The blots were probed with Anti- CBX-5 Recombinant Mouse monoclonal Antibody (Product # 730046, 2 µg/mL) and detected by chemiluminescence Biotin conjugated streptavidin Poly HRP (Product # 21140, 1:10000 dilution). A 22kDa band corresponding to CBX-5 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0341BOX), 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).
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of CBX5 was achieved by transfecting HeLa cells with CBX5 specific validated siRNA (Silencer® select Product # s23885). Western blot analysis was performed using whole cell lysates from the CBX5 knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti- CBX5 Recombinant Mouse monoclonal Antibody (Product # 730046, 2 µg/mL) and Biotin conjugated streptavidin Poly HRP (Product # 21140, 1:10000 dilution). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to CBX5.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of CBX5 on Hela and 293 cells using Anti-CBX5 Recombinant Mouse Monoclonal Antibody (Product # 730046). Hela Cells were fixed with 3% Formaldehyde and lysed with 1% TritonX-100. Cells were then labeled with human Fab and detected with Goat-anti-human Alexa Fluor dye F488 (green). FLAG Cells were double labeled with human Fab and detected with Goat-anti-human Alexa Fluor dye F488 (green) as well as with Mouse-anti-FLAG-M2 and detected with Goat-anti-Mouse Alexa Fluor dye F647 (yellow). The DNA in the nucleus was detected with DAPI (Blue). Control means cells were labeled only with secondary antibody Alexa Fluor Dye.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of CBX5 on Hela and 293 cells using Anti-CBX5 Recombinant Mouse Monoclonal Antibody (Product # 730046). Hela Cells were fixed with 3% Formaldehyde and lysed with 1% TritonX-100. Cells were then labeled with human Fab and detected with Goat-anti-human Alexa Fluor dye F488 (green). FLAG Cells were double labeled with human Fab and detected with Goat-anti-human Alexa Fluor dye F488 (green) as well as with Mouse-anti-FLAG-M2 and detected with Goat-anti-Mouse Alexa Fluor dye F647 (yellow). The DNA in the nucleus was detected with DAPI (Blue). Control means cells were labeled only with secondary antibody Alexa Fluor Dye.
Conjugate: Biotin

730046

Antibody data