Recombinant
RabMAb

Anti-MCL1 antibody [Y37] (ab32087)

Overview

  • Product name
    Anti-MCL1 antibody [Y37]
    See all MCL1 primary antibodies
  • Description
    Rabbit monoclonal [Y37] to MCL1
  • Host species
    Rabbit
  • Specificity
    ab32087 recognises MCL1. The antibody does not cross-react with other Bcl-2 family members.
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-P, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human MCL1 aa 100-200. The exact sequence is proprietary.
    Database link: Q07820
    (Peptide available as ab199979)

  • Positive control
    • WB: A431 cell lysate. ICC/IF: HCT116, MCF7 and H1299 cells. Flow Cyt: A431 cells.
  • General notes

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab32087 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/100 - 1/500.
WB 1/1000 - 1/5000. Predicted molecular weight: 37 kDa.Can be blocked with MCL1 peptide (ab199979).

 

IHC-P 1/100.

See IHC antigen retrieval protocols.

Flow Cyt 1/250.

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.

IP 1/80 - 1/100.

Target

  • Function
    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.
  • Sequence similarities
    Belongs to the Bcl-2 family.
  • Post-translational
    modifications
    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.
  • Cellular localization
    Membrane. Cytoplasm. Mitochondrion. Nucleus > nucleoplasm. Cytoplasmic, associated with mitochondria.
  • Information by UniProt
  • Database links
  • Alternative names
    • Bcl 2 related protein EAT/mcl1 antibody
    • Bcl-2-like protein 3 antibody
    • Bcl-2-related protein EAT/mcl1 antibody
    • BCL2 related antibody
    • Bcl2-L-3 antibody
    • BCL2L3 antibody
    • EAT antibody
    • Induced myeloid leukemia cell differentiation protein Mcl 1 antibody
    • Induced myeloid leukemia cell differentiation protein Mcl-1 antibody
    • MCL 1 antibody
    • MCL1 antibody
    • MCL1-ES antibody
    • mcl1/EAT antibody
    • MCL1_HUMAN antibody
    • MCL1L antibody
    • MCL1S antibody
    • MGC104264 antibody
    • MGC1839 antibody
    • Myeloid Cell Leukemia 1 antibody
    • Myeloid cell leukemia ES antibody
    • Myeloid cell leukemia sequence 1 antibody
    • Myeloid cell leukemia sequence 1 BCL2 related antibody
    • Myeloid cell leukemia sequence 1 isoform 1 antibody
    • OTTHUMP00000032794 antibody
    • OTTHUMP00000032795 antibody
    • TM antibody
    see all

Images

  • All lanes : Anti-MCL1 antibody [Y37] (ab32087) at 1/1000 dilution

    Lane 1 : Human lung tissue with NFDM/TBST
    Lane 2 : Human lung cancer tissue with NFDM/TBST
    Lane 3 : Human liver tissue 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 for samples 1-3: 15 seconds; exposure time for samples 4-6: 1 minute.

    Additional bands: We are unsure as to the identity of these extra bands.

  • 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).

  • 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 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.

    See Abreview

  • 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.

References

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 Dis 9:137 (2018). Read more (PubMed: 29374168) »
  • Ma L  et al. Long non-coding RNA XIST promotes cell growth and invasion through regulating miR-497/MACC1 axis in gastric cancer. Oncotarget 8:4125-4135 (2017). WB ; Human . Read more (PubMed: 27911852) »

See all 42 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Glioblastoma primary cells)
Loading amount
30 µg
Specification
Glioblastoma primary cells
Gel Running Conditions
Reduced Denaturing (gradient gel 5-15%)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Nov 21 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse Embryonic Fibroblast)
Loading amount
20 µg
Specification
Mouse Embryonic Fibroblast
Gel Running Conditions
Reduced Denaturing (10% Bis Bris)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Dec 31 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK 293T)
Loading amount
20 µg
Specification
HEK 293T
Gel Running Conditions
Reduced Denaturing (10% Bis Tris)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Dec 30 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - other (HELA)
Total protein in input
300 µg
Specification
HELA
Immuno-precipitation step
Protein A
Username

Abcam user community

Verified customer

Submitted Nov 29 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cultured Cells (H1299)
Specification
H1299
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton-x
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Username

Abcam user community

Verified customer

Submitted Nov 25 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (HELA)
Loading amount
30 µg
Specification
HELA
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Abcam user community

Verified customer

Submitted Nov 25 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Primary keratinocytes)
Loading amount
30 µg
Specification
Primary keratinocytes
Treatment
15mJ/cm2 UVB
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Dr. Christine Tomlins

Verified customer

Submitted Oct 29 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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