Anti-Bak antibody (ab92999)
Key features and details
- Rabbit polyclonal to Bak
- Suitable for: ICC/IF, WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG
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 & Species
See all applications and species dataApplication Species ICC/IF HumanWB Human
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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 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. 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
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch 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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KO cell pellets
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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.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanWB HumanApplication Abreviews Notes 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).Notes 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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Western blot - Anti-Bak antibody (ab92999)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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Immunocytochemistry/ Immunofluorescence - Anti-Bak antibody (ab92999)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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Western blot - Anti-Bak antibody (ab92999)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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Western blot - Anti-Bak antibody (ab92999)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.
Images
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Western blot - Anti-Bak antibody (ab92999)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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Immunocytochemistry/ Immunofluorescence - Anti-Bak antibody (ab92999)
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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Western blot - Anti-Bak antibody (ab92999)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.
-
Western blot - Anti-Bak antibody (ab92999)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
