Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
For unpurified use 1 ug for 106 cells. (For lot-specific stock concentration, please contact Abcam). ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
1/80 - 1/100.
Involved in the regulation of apoptosis versus cell survival, and in the maintenance of viability but not of proliferation. Mediates its effects by interactions with a number of other regulators of apoptosis. Isoform 1 inhibits apoptosis. Isoform 2 promotes apoptosis.
Belongs to the Bcl-2 family.
Cleaved by CASP3 during apoptosis. In intact cells cleavage occurs preferentially after Asp-127, yielding a pro-apoptotic 28 kDa C-terminal fragment. Rapidly degraded in the absence of phosphorylation on Thr-163 in the PEST region. Phosphorylated on Thr-163. Treatment with taxol or okadaic acid induces phosphorylation on additional sites.
Membrane. Cytoplasm. Mitochondrion. Nucleus > nucleoplasm. Cytoplasmic, associated with mitochondria.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon adenocarcinoma tissue labelling MCL1 with purified ab32087 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Flow Cytometry analysis of RAMOS cells labelling MCL1 with purified ab32087 at 1/250 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
Immunocytochemistry/Immunofluorescence analysis of HCT 116 (human colorectal carcinoma) cells labelling MCL1 with purified ab32087 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).
Western blot - Anti-MCL1 antibody [Y37] (ab32087)
All lanes : Anti-MCL1 antibody [Y37] (ab32087) at 1/1000 dilution
Lane 1 : Ramos cell lysate with NFDM/TBST Lane 2 : HL-60 cell lysate with NFDM/TBST Lane 3 : A431 cell lysate with NFDM/TBST Lane 4 : HeLa cell lysate with NFDM/TBST Lane 5 : MCF-7 cell lysate with NFDM/TBST Lane 6 : HepG2 cell lysate with NFDM/TBST
Lysates/proteins at 20 µg per lane.
Blocking peptides at 5 % per lane.
Secondary All lanes : Goat anti-rabbit IgG (H+L), peroxidase conjugated. at 1/1000 dilution
Predicted band size: 37 kDa
Exposure time: 15 seconds
Additional bands: We are unsure as to the identity of these extra bands.
Immunocytochemistry/ Immunofluorescence - Anti-MCL1 antibody [Y37] (ab32087)This image is courtesy of an anonymous Abreview.
Immunocytochemistry/Immunofluorescence analysis of H1299 cells labelling MCL1 with unpurified ab32087. Cells were PFA-fixed and permeabilized in 0.5% Triton X-100 prior to blocking in 3% Serum for 1 hour at 24°C. The primary antibody was diluted 1/100 and incubated with the sample for 1 hour at 24°C. The secondary antibody was an Alexa Fluor® 488-conjugated Goat anti-Rabbit polyclonal, diluted 1/2000. DAPI (blue) was used as the nuclear counterstain.
Flow Cytometry analysis of A431 cells labelling MCL1 with unpurified ab32087 (red line). Cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32087, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C.
Black - Isotype control, rabbit monoclonal IgG.
Acquisition of >5,000 events was performed. This antibody gave a decreased signal in A431 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.
Immunocytochemistry/Immunofluorescence analysis of HCT116 cells treated with wogonin (ab142471) labelling MCL1 with unpurified ab32087. Decrease of MCL1 expression correlates with increased concentration of wogonin, as described in literature. Cells were incubated at 37°C for 2h in media containing different concentrations of ab142471 (wogonin) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab32087 (1/100) dilution was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
This product has been referenced in:
Li H et al. Downregulation of MCL-1 and upregulation of PUMA using mTOR inhibitors enhance antitumor efficacy of BH3 mimetics in triple-negative breast cancer. Cell Death Dis9:137 (2018).
Read more (PubMed: 29374168) »
Lin J et al. FBW7 is associated with prognosis, inhibits malignancies and enhances temozolomide sensitivity in glioblastoma cells. Cancer Sci109:1001-1011 (2018).
Read more (PubMed: 29427543) »