Recombinant
RabMAb

Recombinant Anti-MCL1 antibody [Y37] - BSA and Azide free (ab186822)

Overview

  • Product name

    Anti-MCL1 antibody [Y37] - BSA and Azide free
    See all MCL1 primary antibodies
  • Description

    Rabbit monoclonal [Y37] to MCL1 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, IP, ICC/IF, Flow Cyt, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human MCL1 aa 100-200. The exact sequence is proprietary.

  • Positive control

    • A431 cell lysate, cervical carcinoma, human tonsil.
  • General notes

    Ab186822 is the carrier-free version of ab32087. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab186822 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab186822 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
IHC-P Use at an assay dependent concentration.

See IHC antigen retrieval protocols.

IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

WB Use at an assay dependent concentration. Predicted molecular weight: 37 kDa.

Can be blocked with MCL1 peptide (ab199979).

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

  • This IHC data was generated using the same anti-MCL1 antibody clone, Y37, in a different buffer formulation (cat# ab32087).

    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 (human Burkitt's lymphoma cell line) 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32087).

  • Immunocytochemistry/Immunofluorescence analysis of HCT 116 (human colorectal carcinoma cell line) 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32087).

  • Immunocytochemistry/Immunofluorescence analysis of HCT 116 (human colorectal carcinoma cell line) 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 data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32087).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32087).

  • Flow Cytometry analysis of A431 (human epidermoid carcinoma cell line) 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32087).

  • This ICC/IF data was generated using the same anti-MCL1 antibody clone, Y37, in a different buffer formulation (cat# ab32087).

    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.

References

This product has been referenced in:

  • Cippà PE  et al. Targeting apoptosis to induce stable mixed hematopoietic chimerism and long-term allograft survival without myelosuppressive conditioning in mice. Blood 122:1669-77 (2013). Read more (PubMed: 23869083) »
  • Yallapu MM  et al. Novel curcumin-loaded magnetic nanoparticles for pancreatic cancer treatment. Mol Cancer Ther 12:1471-80 (2013). Read more (PubMed: 23704793) »
See all 13 Publications for this product

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
No
Specification
HeLa
Fixative
Methanol

Dr. Kirk Mcmanus

Verified customer

Submitted Jul 03 2018

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