重组
RabMAb

Anti-MCL1抗体[Y37] (ab32087)

概述

  • 产品名称
    Anti-MCL1抗体[Y37]
    参阅全部 MCL1 一抗
  • 描述
    兔单克隆抗体[Y37] to MCL1
  • 特异性
    ab32087 recognises MCL1. The antibody does not cross-react with other Bcl-2 family members.
  • 经测试应用
    适用于: ICC/IF, WB, IHC-P, Flow Cyt, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human MCL1 aa 100-200.
    Database link: Q07820
    (Peptide available as ab199979)

  • 阳性对照
    • WB: A431 cell lysate. ICC/IF: HCT116, MCF7 and H1299 cells. Flow Cyt: A431 cells.
  • 常规说明

    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.

性能

应用

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.

应用 Ab评论 说明
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 protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

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.

靶标

  • 功能
    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.
  • 序列相似性
    Belongs to the Bcl-2 family.
  • 翻译后修饰
    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.
  • 细胞定位
    Membrane. Cytoplasm. Mitochondrion. Nucleus > nucleoplasm. Cytoplasmic, associated with mitochondria.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

图片

  • 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
    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
    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.

文献

This product has been referenced in:
  • 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) »
  • Peperzak V  et al. Functional disparities among BCL-2 members in tonsillar and leukemic B-cell subsets assessed by BH3-mimetic profiling. Cell Death Differ 24:111-119 (2017). Read more (PubMed: 27689871) »

See all 34 Publications for this product

客户评价及客户问答




I can confirm that we have not tested if this antibody is also binding to MCL1 in complexes.


I would like to point out that the clone [Y37] (parent product ab32087) is tested for IP. We have customer feedback where this clo...

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I can confirm that the immunogen sequence for ab32087 has high alignment to both isoforms 1 and 2 from the following Uniprot protein database page for the protein:
http://www.uniprot.org/uniprot/Q07820

It is therefore very likely to det...

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Thank you for contacting us.

The concentration of lot GR87086-2 is 0.81 mg/ml.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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

提交于 Nov 21 2012

Gracias por contactar con Abcam.

Mi compañera no se encuentra hoy en la oficina.

Volvera a estar disponible el lunes 12 de noviembre.

Para cuestiones urgentes, puede ponerse en contacto con mis compañeros en ...

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Thank you for contacting us.

I am sorry to hear you have been having some difficulty with the lot number GR42511-4 of the anti-MCL1 antibody [Y37] (ab32087). Following your concern I have looked back into our previous records and we have as y...

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Thank you for your enquiry.

I can confirm this antibody has been tested for flow cytometry at dilution 1:50. The IgG concentration of the original lot GR42511-5 is 0.235 mg/mL.

I hope this will be helpful. Please let me know if yo...

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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

提交于 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

提交于 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

提交于 Nov 29 2010

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

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