Anti-Bak antibody (ab92999)
Key features and details
- Rabbit polyclonal to Bak
- Suitable for: ICC/IF, WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
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- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-Bak antibody
See all Bak primary antibodies -
Description
Rabbit polyclonal to Bak -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide corresponding to Human Bak aa 1-100 conjugated to keyhole limpet haemocyanin.
(Peptide available asab106421) -
Positive control
- WB: HeLa, HAP1 and A431 cell lysates. ICC/IF: HepG2 cells.
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General notes
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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab92999 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 |
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ICC/IF |
Use a concentration of 5 µg/ml.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 25 kDa (predicted molecular weight: 23 kDa).
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Notes |
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ICC/IF
Use a concentration of 5 µg/ml. |
WB
Use at an assay dependent concentration. Detects a band of approximately 25 kDa (predicted molecular weight: 23 kDa). |
Target
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Function
In the presence of an appropriate stimulus, accelerates programmed cell death by binding to, and antagonizing the anti-apoptotic action of BCL2 or its adenovirus homolog E1B 19k protein. Low micromolar levels of zinc ions inhibit the promotion of apoptosis. -
Tissue specificity
Expressed in a wide variety of tissues, with highest levels in the heart and skeletal muscle. -
Sequence similarities
Belongs to the Bcl-2 family. -
Domain
Intact BH3 motif is required by BIK, BID, BAK, BAD and BAX for their pro-apoptotic activity and for their interaction with anti-apoptotic members of the Bcl-2 family. -
Cellular localization
Mitochondrion membrane. - Information by UniProt
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Database links
- Entrez Gene: 578 Human
- Omim: 600516 Human
- SwissProt: Q16611 Human
- Unigene: 485139 Human
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Alternative names
- Apoptosis regulator BAK antibody
- BAK antibody
- BAK like antibody
see all
Images
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All lanes : Anti-Bak antibody (ab92999) at 1 µg/ml
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : BAK1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 23 kDaLanes 1- 2: Merged signal (red and green). Green - ab92999 observed at 23 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab92999 was shown to react with Bak in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265277 (knockout cell lysate ab257077) was used. Wild-type HeLa and BAK1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab92999 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 µg/ml and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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ICC/IF image of ab92999 stained HepG2 cells. The cells were 4% formaldehyde fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab92999 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit IgG (H+L)(ab96899) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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All lanes : Anti-Bak antibody (ab92999) at 1 µg/ml
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Bak knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 23 kDaLanes 1 - 2: Merged signal (red and green). Green - ab92999 observed at 23 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab92999 was shown to recognize Bak in wild-type HAP1 cells as signal was lost at the expected MW in Bak knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Bak knockout samples were subjected to SDS-PAGE. ab92999 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Anti-Bak antibody (ab92999) at 1 µg/ml + A431 (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?
Additional bands at: 70 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 12 minutes
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab92999 has not yet been referenced specifically in any publications.