Recombinant Anti-Bim antibody [Y36] (ab32158)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y36] to Bim
- Suitable for: WB, IHC-P, Flow Cyt, IP, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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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 & Species
Application Species Flow Cyt HumanICC/IF MouseHumanIHC-P HumanIP HumanWB MouseRatHuman -
Immunogen
Synthetic peptide within Human Bim aa 1-100. The exact sequence is proprietary.
(Peptide available asab179844) -
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.
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General notes
Mouse and Rat species are recommended based on WB results, we do not guarantee IHC-P for Mouse and Rat.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
IgG fraction -
Clonality
Monoclonal -
Clone number
Y36 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
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Flow Cyt |
Human
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ICC/IF |
Mouse
Human
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IHC-P |
Human
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IP |
Human
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WB |
Mouse
Rat
Human
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Application | Abreviews | Notes |
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WB | (5) |
1/500 - 1/2000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).Can be blocked with Bim peptide (ab179844).
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IHC-P |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. Mouse and Rat species are recommended based on WB results, we do not guarantee IHC-P for Mouse and Rat. |
|
Flow Cyt |
1/50 - 1/100.
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IP | (1) |
1/40 - 1/50.
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ICC/IF |
1/100 - 1/250.
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Notes |
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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. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. Mouse and Rat species are recommended based on WB results, we do not guarantee IHC-P for Mouse and Rat. |
Flow Cyt
1/50 - 1/100. |
IP
1/40 - 1/50. |
ICC/IF
1/100 - 1/250. |
Target
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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
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Database links
- Entrez Gene: 10018 Human
- Entrez Gene: 12125 Mouse
- Entrez Gene: 64547 Rat
- Omim: 603827 Human
- SwissProt: O43521 Human
- SwissProt: O54918 Mouse
- SwissProt: O88498 Rat
- Unigene: 469658 Human
see all -
Alternative names
- BCL2 like 11 antibody
- B2L11_HUMAN antibody
- BAM antibody
see all
Images
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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.
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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. -
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
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bim antibody [Y36] (ab32158)
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.
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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.
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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 Detection Reagent (HRP) was used for detection (1/10 000).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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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 kDaObserved 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)
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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
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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.
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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 kDaObserved 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)
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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 secondsPrimary diluted in PBS (5% BSA + 0.1% tween20) and incubated with sample for 1 hour and 30 minutes at 20°C.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (61)
ab32158 has been referenced in 61 publications.
- Feng C et al. Neuroprotective Effect of Danhong Injection on Cerebral Ischemia-Reperfusion Injury in Rats by Activation of the PI3K-Akt Pathway. Front Pharmacol 11:298 (2020). PubMed: 32218735
- Kabir S et al. The CUL5 ubiquitin ligase complex mediates resistance to CDK9 and MCL1 inhibitors in lung cancer cells. Elife 8:N/A (2019). PubMed: 31294695
- Yuan F et al. A New Regulatory Mechanism Between P53 And YAP Crosstalk By SIRT1 Mediated Deacetylation To Regulate Cell Cycle And Apoptosis In A549 Cell Lines. Cancer Manag Res 11:8619-8633 (2019). PubMed: 31576168
- Gao K et al. Crocetin protects against fulminant hepatic failure induced by lipopolysaccharide/D-galactosamine by decreasing apoptosis, inflammation and oxidative stress in a rat model. Exp Ther Med 18:3775-3782 (2019). PubMed: 31616509
- Huang Q et al. Effect of baicalin on proliferation and apoptosis in pancreatic cancer cells. Am J Transl Res 11:5645-5654 (2019). PubMed: 31632536