Anti Mcl-1  

Immunocytochemistry of Mcl-1 antibody. ？ Cell Type: ？Raji cells. ？ Fixation: ？formalin fixed paraffin embedded. ？ Antigen retrieval: ？not required. Primary antibody: ？Mcl-1 antibody at 10 μg/mL ？for 1 h at RT. Secondary antibody: ？Peroxidase rabbit secondary antibody at 1:10,000 for 45 min at RT. Localization: ？Mcl-1 is located in the cytoplasm, cell membrane, mitochondrion, and the nucleus. Staining: ？Mcl-1 is stained brown with hematoxylin purple counterstain.

Immunofluorescence Microscopy of Mcl-1 antibody. ？ Cell Type: ？Raji cells. ？ Fixation: ？0.5% PFA.？ Antigen retrieval: ？not required. Primary antibody: ？Mcl-1 antibody ？at 20 μg/mL ？for 1 h at RT. Secondary antibody: ？Fluorescein rabbit secondary antibody at 1:10,000 for 45 min at RT. Localization: ？Mcl-1 is located in the cytoplasm, cell membrane, mitochondrion, and the nuclues. Staining: ？Mcl-1 as red fluorescent signal.

Western Blot of Mcl-1 antibody in Raji cell lysates. ？ Lane A: Mcl-1 antibody at 1 μg/mL. ？ Lane B: Mcl-1 antibody at 2 μg/mL. ？ Load: ？35 μg per lane. Primary antibody: ？Mcl-1 antibody at designated concentrations for overnight at 4°C. Secondary antibody: ？ Peroxidase rabbit secondary antibody at 1:10,000 for 45 min at RT. Block: ？5% BLOTTO overnight at 4°C. Predicted/Observed size: ？37 kDa, 42 kDa for Mcl-1.？ Other band(s): Mcl-1 splice variants and isoforms.

A.？PC3, H1299, and Reh ABT-R cells were transfected with siCyclin E and siControl and treated with ABT-737 (10 μM) after 24 h. Expression levels of the indicated proteins were determined by immunoblotting after an additional 18 h of incubation. Reh ABT-R cells were treated with dinaciclib for 24 h and？B.？expression levels for the indicated proteins was determined by immunoblotting？C.？Mcl-1 and？D.？Bim mRNA was analyzed by qRT-PCR.？E.？Reh ABT-R cells were treated with dinaciclib (10 nM) for 18 h. Association of Mcl-1 with Bim was determined by immunoblotting in Reh and Reh ABT-R cells by immunoprecipitating Mcl-1.？F.？Association of Bim with Mcl-1 was determined by immunoprecipitating Bim in Reh ABT-R cells after treating with dinaciclib for 18 h.？G.？DOHH2 and？H.？SU-DHL-16？I.？Reh ABT-R cells were treated with the indicated concentrations of ABT-199 ± dinaciclib for 24 h. Percentage reduction in metabolic activity was determined by the MTS assay.？J.？Combination index (CI) values for indicated cell lines. CI< 1 indicates synergism. Data in？A.,？B.,？E.,？F.？are representative of three independent experiments. SD in？C.,？D.,？G.-I.？is indicated by error bars (n？= 3). Fig 6. PMID:？26219338