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

  • Western blot analysis of PUMA expression in human (H) K562 and mouse (M) 3T3 cell lysates with anti-PUMA at 2 µg /ml.

    Western blot analysis of PUMA expression in human (H) K562 and mouse (M) 3T3 cell lysates with anti-PUMA at 2 µg /ml.
  • Ab9643 staining human ovarian carcinoma tissue. Staining is localized to mitochondria.
    Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • 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) »
  • Holmberg Olausson K  et al. Loss of nucleolar histone chaperone NPM1 triggers rearrangement of heterochromatin and synergizes with a deficiency in DNA methyltransferase DNMT3A to drive ribosomal DNA transcription. J Biol Chem 289:34601-19 (2014). Read more (PubMed: 25349213) »

See all 43 Publications for this product

Product Wall

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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Verified customer

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

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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

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (brain)
Loading amount 25 µg
Specification brain
Treatment saline 0.9%-6 hrs, ethanol 20%-6 hrs
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C
Username

Arindam Ghosh

Verified customer

Submitted Mar 08 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (Mouse Embryonic Fibroblasts)
Loading amount 30 µg
Specification Mouse Embryonic Fibroblasts
Gel Running Conditions Reduced Denaturing (10% Bis Tris)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Mr. Brian Koss

Verified customer

Submitted Jan 06 2011

1-10 of 17 Abreviews or Q&A

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