Validated using a knockout cell line
Recombinant
RabMAb

Anti-Bim antibody [Y36] (ab32158)

Overview

  • Product name
    Anti-Bim antibody [Y36]
    See all Bim primary antibodies
  • Description
    Rabbit monoclonal [Y36] to Bim
  • Host species
    Rabbit
  • Specificity
    The antibody can recognize all isoforms of Bim. The antibody does not cross-react with other Bcl-2 family members.
  • Tested applications
    Suitable for: IHC-FoFr, WB, IHC-P, Flow Cyt, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Bim aa 1-100. The exact sequence is proprietary.
    (Peptide available as ab179844)

  • Positive control
    • WB : Raji, Jurkat, A431, Molt-4, A20, MEF, Raw264.7 and PC-12 cell lysate; Human and mouse thymus, mouse and rat spleen tissue lysate. IHC : breast carcinoma tissue. ICC/IF : A20 and Raji cells. FC : A431 and Raji whole cell lysate. HAP1-WT cells. IP : Raji whole cell lysate.
  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    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.

Applications

Our Abpromise guarantee covers the use of ab32158 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-FoFr 1/100.
WB 1/500 - 1/2000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).Can be blocked with Bim peptide (ab179844).
IHC-P 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Flow Cyt 1/50 - 1/100.
IP 1/40 - 1/50.
ICC/IF 1/100 - 1/250.

Target

  • Function
    Induces apoptosis. Isoform BimL is more potent than isoform BimEL. Isoform Bim-alpha1, isoform Bim-alpha2 and isoform Bim-alpha3 induce apoptosis, although less potent than the isoforms BimEL, BimL and BimS. Isoform Bim-gamma induces apoptosis.
  • Tissue specificity
    Isoform BimEL, isoform BimL and isoform BimS are the predominant isoforms and are ubiquitously expressed with a tissue-specific variation. Isoform Bim-gamma is most abundantly expressed in small intestine and colon, and in lower levels in spleen, prostate, testis, heart, liver and kidney.
  • Sequence similarities
    Belongs to the Bcl-2 family.
  • Domain
    The BH3 motif is required for Bcl-2 binding and cytotoxicity.
  • Cellular localization
    Mitochondrion and Endomembrane system. Associated with intracytoplasmic membranes.
  • Information by UniProt
  • Database links
  • Alternative names
    • BCL2 like 11 antibody
    • B2L11_HUMAN antibody
    • BAM antibody
    • Bcl 2 interacting protein Bim antibody
    • Bcl 2 related ovarian death agonist antibody
    • Bcl-2-like protein 11 antibody
    • BCL2 interacting mediator of cell death antibody
    • BCL2 like 11 (apoptosis facilitator) antibody
    • BCL2 like protein 11 antibody
    • Bcl2-interacting mediator of cell death antibody
    • Bcl2-L-11 antibody
    • Bcl2l11 antibody
    • BIM alpha6 antibody
    • BIM antibody
    • BIM beta6 antibody
    • BIM beta7 antibody
    • BimEL antibody
    • BimL antibody
    • BOD antibody
    see all

Images

  • Overlay histogram showing HAP1 wildtype (green line) and HAP1-BCL2L11 knockout cells (red line) stained with ab32158. The cells were fixed 80% methanol (5 min), and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab32158, 1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) presorbed (ab150081) at 1/2000 dilution for 30 min at 22°C.

    A rabbit IgG1 isotype control antibody  (ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-BCL2L11  knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

    Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

  • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: Bim knockout HAP1 whole cell lysate (20 µg)
    Lane 3: Raji whole cell lysate (20 µg)
    Lane 4: Jurkat whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab32158 observed at 25 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab32158 was shown to specifically react with Bim when Bim knockout samples were used. Wild-type and Bim knockout samples were subjected to SDS-PAGE. ab32158 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/Immunofluorescence analysis of Raji (Human Burkitt's lymphoma cell line) labeling Bim with ab32158 at a dilution of 1/250. Cells were fixed with 100% methanol. Ab150077 (1/1000) was used as the secondary antibody. Cells were co-stained with ab7291, a mouse anti-tubulin antibody (1/200) using ab150120 as the secondary. Nuclei were counterstained with DAPI (blue).

    Secondary antibody only control, cells without incubation with the primary antibody was used as negative control.

    Confocal image showing cytoplasmic staining on Raji cell line

  • ab32158 staining Bim in human breast cancer tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Antigen retrieval was by heat mediated antigen retrieval using Tris/EDTA Buffer, PH9 (ab93684). Samples were incubated with primary antibody (1/100 in blocking buffer) and a Biotin-conjugated Donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody. Cytoplasmic staining can be seen in the human breast cancer cells. Hematoxylin was used as a counter stain.

  • Flow Cytometry analysis of Raji (human Burkitt's lymphoma) whole cell lysate labeling Bim with ab32158 at 1/100 (red). Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488)-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG (ab172730). Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Ab32158 at 1/50 immunoprecipitating Bim in Raji (human Burkitt's lymphoma) whole cell lysate.

    Lane 1 (input): Raji whole cell lysate (10µg)

    Lane 2 (+): ab32158 + Raji whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32158 in Raji whole cell lysate.

    For western blotting, ab32158 (1/1000) was used as the primary antibody and ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/10 000).

     

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-Bim antibody [Y36] (ab32158) at 1/2000 dilution

    Lane 1 : Raji (human Burkitt's lymphoma) whole cell lysate
    Lane 2 : A431 (human epidermoid carcinoma) whole cell lysate
    Lane 3 : Molt-4 (human acute lymphoblastic leukemia) whole cell lysate
    Lane 4 : Human thymus tissue lysate
    Lane 5 : Mouse thymus tissue lysate
    Lane 6 : A20 (mouse reticulum cell sarcoma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 22 kDa



    Observed band size : 22, 18 kDa

    Exposure time : Lane 1- 5: 3 minutes; Lane 6: 2 seconds

    Blocking/Diluting buffer and concentration : 5% NFDM /TBST 

    The observed molecular weight is consistent with the literature (PMID: 24872388)

  • Immunocytochemistry/Immunofluorescence analysis of A20 (Mouse reticulum sarcoma cell line) labeling Bim with ab32158 at a dilution of 1/250. Cells were fixed with 100% methanol. Ab150077 (1/1000) was used as the secondary antibody. Cells were co-stained with ab7291, a mouse anti-tubulin antibody (1/200) using ab150120 as the secondary. Nuclei were counterstained with DAPI (blue). 

    Secondary antibody only control, cells without incubation with the primary antibody was used as negative control.

    Confocal image showing cytoplasmic staining on A20 cell line

  • Flow Cytometry analysis of A431 (human epidermoid carcinoma) cells labelling Bim with ab32158 at 1/50 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • All lanes : Anti-Bim antibody [Y36] (ab32158) at 1/2000 dilution

    Lane 1 : Mouse spleen tissue lysate
    Lane 2 : Rat spleen tissue lysate
    Lane 3 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 4 : Raw264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 22 kDa



    Observed band size : 22, 18 kDa

    Exposure time : Lane 1-3: 3 minutes; Lane 4: 10 seconds

    Blocking/Diluting buffer and concentration : 5% NFDM /TBST

    The observed molecular weight is consistent with the literature (PMID: 24872388)

  • Anti-Bim antibody [Y36] (ab32158) at 1/500 dilution + Whole cell lysates prepared from human jurkat cells at 200000 cells

    Secondary
    HRP conjugated Donkey polyclonal to rabbit IgG at 1/2000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 22 kDa
    Observed band size: 22 kDa


    Exposure time: 30 seconds


    Primary diluted in PBS (5% BSA + 0.1% tween20) and incubated with sample for 1 hour and 30 minutes at 20°C.

References

This product has been referenced in:
  • Liu B  et al. RACKI induces chemotherapy resistance in esophageal carcinoma by upregulating the PI3K/AKT pathway and Bcl-2 expression. Onco Targets Ther 11:211-220 (2018). Read more (PubMed: 29379302) »
  • Ruvolo PP  et al. Role of MSC-derived galectin 3 in the AML microenvironment. Biochim Biophys Acta 1865:959-969 (2018). Read more (PubMed: 29655803) »

See all 36 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Immuno-precipitation step
Protein A/G
Sample
Human Cell lysate - whole cell (HCT116 p53-/- cell line)
Specification
HCT116 p53-/- cell line
Total protein in input
100 µg
Username

Mr. Christian Marx

Verified customer

Submitted Aug 27 2013

Application
Western blot
Sample
Mouse Cell lysate - whole cell (MEF cell line)
Loading amount
60 µg
Specification
MEF cell line
Treatment
DMSO control and 2 µM SAHA for 24 hrs
Gel Running Conditions
Reduced Denaturing (13%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Christian Marx

Verified customer

Submitted Feb 13 2013

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (HCT116 colon cancer cell line wt and p53 negative)
Loading amount
60 µg
Specification
HCT116 colon cancer cell line wt and p53 negative
Gel Running Conditions
Reduced Denaturing (10)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Christian Marx

Verified customer

Submitted Oct 26 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Jurkat cells)
Loading amount
200000 cells
Specification
Jurkat cells
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Nov 24 2009

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

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