Overview

  • Product nameAnti-PUMA antibody
    See all PUMA primary antibodies
  • Description
    Rabbit polyclonal to PUMA
  • SpecificityAt least 2 isoforms are known to exist; this antibody will detect both isoforms.
  • Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide:

    PLPRGHRAPEMEPN

    , corresponding to C terminal amino acids 180-193 of Human PUMA alpha. (Peptide available as ab9644.)

  • Positive control
    • K562 or 3T3
  • General notes


    Apoptosis is related to many diseases and development. The p53 tumor-suppressor protein induces apoptosis through transcriptional activation of several genes. A novel p53 inducible pro-apoptotic gene was identified recently and designated PUMA (for p53 upregulated modulator of apoptosis) and bbc3 (for Bcl-2 binding component 3) in human and mouse (1-3). PUMA/bbc3 is one of the pro-apoptotic Bcl-2 family members including Bax and Noxa, which are also transcriptional targets of p53. The PUMA gene encodes two BH3 domain-containing proteins termed PUMA-a and PUMA-b (1). PUMA proteins bind Bcl-2, localize to the mitochondria, and induce cytochrome c release and apoptosis in response to p53. PUMA may be a direct mediator of p53-induced apoptosis.

Properties

Applications

Our Abpromise guarantee covers the use of ab9643 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
WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 23 kDa.Can be blocked with Human PUMA peptide (ab9644). Use at a concentration of 1 - 2 µg/ml. Detects a band of approximately 23 kDa. Can be blocked with PUMA peptide (180/193) (ab9644). A lower band at approximately 16 kDa was detected in MOLT4 and U937 cells, which may represent the PUMA-beta form.
IHC-P Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 µg/ml.

Target

Anti-PUMA antibody images

  • Anti-PUMA antibody (ab9643) at 1/1000 dilution + HeLa whole cell lysate

    Secondary
    Alexa Fluor 680-conjugated goat anti-rabbit IgG polyclonal at 1/1 dilution

    Observed band size : 20 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 26 kDa (possible non-specific binding),42 kDa (possible non-specific binding),50 kDa (possible non-specific binding).

    Exposure time : 5 seconds

    This image is courtesy of an anonymous Abreview

    Blocked with 5% milk for 1 hour.

    Incubated with the primary antibody for 18 hours.

    See Abreview

  • ab9643 staining rat gonad tissue sections by Immunohistochemistry (IHC-P - Formalin/PFA-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 1.5% milk for 45 minutes at 25°C. Protease/PBS (100mg/200ml) was used for antigen retrieval, H2O2/methanol solution (0.3%) was used to destroy endogenous peroxidase activity. Samples were incubated with the primary antibody (1/500) for 12 hours at 4°C. A Biotin-conjugated goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.

    See Abreview

  • ICC/IF image of ab9643 stained HeLa cells. The cells were 4% PFA fixed (10 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 (ab9643, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) 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.

References for Anti-PUMA antibody (ab9643)

This product has been referenced in:
  • Liu ZQ  et al. Expression of PUMA in Follicular Granulosa Cells Regulated by FoxO1 Activation During Oxidative Stress. Reprod Sci 22:696-705 (2015). IHC . Read more (PubMed: 25425107) »
  • Singh SP  et al. HIF-1a Plays a Critical Role in the Gestational Sidestream Smoke-Induced Bronchopulmonary Dysplasia in Mice. PLoS One 10:e0137757 (2015). WB ; Mouse . Read more (PubMed: 26361040) »

See all 44 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (human hepatocytes)
Gel Running Conditions Reduced Denaturing
Loading amount 20 µg
Treatment 100 ng/ml for hepatocytes
Specification human hepatocytes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted Apr 24 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (human hepatocytes)
Gel Running Conditions Reduced Denaturing
Loading amount 20 µg
Specification human hepatocytes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted Mar 13 2017

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Gonad)
Antigen retrieval step Enzymatic
Permeabilization No
Specification Gonad
Blocking step Milk as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 25°C
Fixative Paraformaldehyde
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Submitted Apr 14 2015

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (hepatocytes)
Specification hepatocytes
Treatment 100 U/ml IL-1 beta
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted Dec 23 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (10% SDS PAGE)
Sample Mouse Tissue lysate - whole (Liver)
Specification Liver
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Jul 22 2014

Thank you for contacting us.


I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to con...

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Thank you for contacting Abcam.

You are correct that you should be seeing bands at 23 &16kDa. I am wondering if the bands that you are seeing at 56 & 72kDa is due to the protein being in a complex with other proteins.

How lo...

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Thank you for contacting Abcam. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement from a new lot. To check the status of the or...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Liver)
Loading amount 40 µg
Specification Liver
Treatment 2Hrs post 6Gy IR
Gel Running Conditions Reduced Denaturing (12% gel)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Sep 26 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing (12)
Sample Human Cell lysate - whole cell (U87)
Specification U87
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Username

Mr. Sanjib Chaudhary

Verified customer

Submitted Aug 12 2011

1-10 of 19 Abreviews or Q&A

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