Anti-MCL1 antibody (ab28147)

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Overview

  • Product name
    Anti-MCL1 antibody
    See all MCL1 primary antibodies
  • Description
    Rabbit polyclonal to MCL1
  • Host species
    Rabbit
  • Specificity
    This antibody detects an ~42/43 kDa protein doublet, corresponding to the expected molecular mass of Mcl1 on SDS-PAGE immunoblots. The antibody specificity is confirmed by peptide inhibition studies. An additional protein at ~33kDa may also be detected. Other additional proteins may also be seen in some samples.
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Rabbit, Guinea pig, Dog, Human, Pig, Monkey
  • Immunogen

    Synthetic peptide corresponding to Human MCL1 aa 121-139 conjugated to Keyhole Limpet Haemocyanin (KLH).
    Sequence:

    SPEEELDGYEPEPLGKRPA

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control
    • HeLa heat shocked cell lysate IHC-P: FFPE human lung tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab28147 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
WB Use a concentration of 0.1 µg/ml. Predicted molecular weight: 42-43 kDa.

0.1 µg/ml was sufficient for detection of Mcl1 in 20µg of HeLa Heat Shocked Cell Lysate by ECL immunoblot analysis.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.

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

    • Western blot - Anti-MCL1 antibody (ab28147)
      Western blot - Anti-MCL1 antibody (ab28147)
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCL1 antibody (ab28147)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCL1 antibody (ab28147)

      IHC image of MCL1 staining in human lung formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab28147, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    • Immunocytochemistry/ Immunofluorescence - Anti-MCL1 antibody (ab28147)
      Immunocytochemistry/ Immunofluorescence - Anti-MCL1 antibody (ab28147)

      ab28147 staining MCL1 in human Hella cells by Immunocytochemistry/ immunofluorescence.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCL1 antibody (ab28147)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCL1 antibody (ab28147)

      ab28147 at 1/50 dillution staining MCL1 in human breast cancer tissue section by Immunohistochemistry.

    References

    This product has been referenced in:
    • Cui Y  et al. Upregulation of microRNA-383 inhibits the proliferation, migration and invasion of colon cancer cells. Oncol Lett 15:1184-1190 (2018). Read more (PubMed: 29399173) »
    • Loriot Y  et al. Radiosensitization by a novel Bcl-2 and Bcl-XL inhibitor S44563 in small-cell lung cancer. Cell Death Dis 5:e1423 (2014). WB ; Human . Read more (PubMed: 25232677) »
    See all 2 Publications for this product

    Publishing research using ab28147? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Question

    I am attaching the imagine from my immunoblot. Thank you.

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    Answer

    Thank you for your enquiry and for providing me with your immunoblot. The bands that you are detecting appear to very clean and specific. I consider the slight aberration that you are detecting a consequence of the molecular weight markers that you are using. We frequently find that molecular weight markers do not always run with a predictable mobility and electrophoresing two markers of similar size in parallel may give unexpected results with the "heavier" molecular weight marker running faster than the "lighter" marker. The slight shift in size may also be attributable to the species that you are using. The doublet that is described on the antibody datasheet was generated from a human (HeLa cell lysate). I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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    Question

    I am writing in regards to the Mcl-1 antibody. When I used this antibody for immunoblot, I saw a predominant band that is 50kDa and not the 42/43 kDa bands as suggested. Does this antibody recognize at a higher molecular weight, so is the 50 kDa band that I am seeing Mcl-1?

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    Answer

    Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. I would very much appreciate it if you could provide me with a blot highlighting the bands that you are detecting. It is a possibility that that the molecular weight markers that you have been employing are migrating at a rate that gives the impression that the bands you have been detecting are of a higher molecular weight that they actually are. We frequently find that small differences in the predicted and actual molecular weight can be attributed to variability in the migration of many of the commercial molecular weight standards available. I look forward to hearing from you with your comments.

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