Recombinant
RabMAb

Anti-PUMA antibody [EP512Y] (ab33906)

Overview

  • Product name
    Anti-PUMA antibody [EP512Y]
    See all PUMA primary antibodies
  • Description
    Rabbit monoclonal [EP512Y] to PUMA
  • Host species
    Rabbit
  • Specificity
    This antibody is specific for both the alpha and beta isoforms of PUMA. Isoform B is primarily recognized on Western blot gel.
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human PUMA aa 150 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: Q9BXH1
    (Peptide available as ab206123)

  • Positive control
    • K562 cell lysate, human breast carcinoma tissue, human ovary carcinoma tissue and Hela cells.
  • General notes

    Mouse: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab33906 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
Flow Cyt 1/30 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB 1/1000 - 1/5000. Detects a band of approximately 18-24 kDa (predicted molecular weight: 21 kDa).Can be blocked with PUMA peptide (ab206123).
IHC-P Use at an assay dependent concentration.
ICC/IF 1/250 - 1/500.

Target

  • Function
    Essential mediator of p53-dependent and p53-independent apoptosis.
  • Tissue specificity
    Ubiquitously expressed.
  • Sequence similarities
    Belongs to the Bcl-2 family.
  • Cellular localization
    Mitochondrion. Localized to the mitochondria in order to induce cytochrome c release.
  • Information by UniProt
  • Database links
  • Alternative names
    • BBC 3 antibody
    • Bbc3 antibody
    • BBC3_HUMAN antibody
    • BCL 2 binding component 3 antibody
    • Bcl-2-binding component 3 antibody
    • BCL2 binding component 3 antibody
    • JFY 1 antibody
    • JFY-1 antibody
    • JFY1 antibody
    • p53 up regulated modulator of apoptosis antibody
    • p53 up-regulated modulator of apoptosis antibody
    • p53 Upregulated Modulator of Apoptosis antibody
    • PUMA alpha antibody
    • PUMA/JFY1 antibody
    see all

Images

  • ab33906 at a 1:250 dilution staining PUMA in Hela cells using Immunocytochemistry/Immunofluorescence.
  • ab33906 at a 1:100 dilution staining PUMA in human ovary carcinoma using Immunohistochemistry, Paraffin Embedded Tissue.
  • Anti-PUMA antibody [EP512Y] (ab33906) at 1/5000 dilution + K562 cell lysate.

    Predicted band size: 21 kDa
    Observed band size: 18 kDa (why is the actual band size different from the predicted?)

  • ab33906 at a 1:100 dilution staining PUMA in human breast carcinoma using Immunohistochemistry, Paraffin Embedded Tissue.
  • Overlay histogram showing K562 cells stained with ab33906 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab33906, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in K562 cells fixed with methanol (5 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.

References

This product has been referenced in:
  • Wang Y  et al. PAK1 overexpression promotes cell proliferation in cutaneous T cell lymphoma via suppression of PUMA and p21. J Dermatol Sci N/A:N/A (2018). WB . Read more (PubMed: 29307600) »
  • Yokoyama T  et al. Apoptosis is augmented in high-grade serous ovarian cancer by the combined inhibition of Bcl-2/Bcl-xL and PARP. Int J Oncol N/A:N/A (2017). WB ; Human . Read more (PubMed: 28350129) »

See all 20 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (paraffin-embedded Human prostate tissue)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 6.0 Citric Acid
Permeabilization
Yes - Tween-20
Specification
paraffin-embedded Human prostate tissue
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 09 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (293T, K562, MCF7)
Gel Running Conditions
Reduced Denaturing (15% SDS-PAGE)
Loading amount
15 µg
Specification
293T, K562, MCF7
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jan 09 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - Tween-20
Specification
HeLa
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Acetone
Username

Abcam user community

Verified customer

Submitted Jan 09 2018

Application
Immunoprecipitation
Sample
Human Cell lysate - other (Mec1 (lymphocytic chronic leukemia))
Total protein in input
2e+007 cells
Immuno-precipitation step
Protein G
Specification
Mec1 (lymphocytic chronic leukemia)
Username

Mrs. Isabel Marzo

Verified customer

Submitted Apr 18 2016

ab33906 lot YH090207C - 0.3260 mg/ml

ab33862 lot YH082410D - 0.6380 mg/ml

ab32429 lot YG080503 - 0.0790 mg/ml

ab40660 lot YI070802CS - 0.1030 mg/ml

ab32445 lot YI072804DS - 0.0250 mg/ml

Thank you for your enquiry.

1) The concentration for the lot YH090207C is 0.3260mg/ml.

2) The lab typically uses 1XPBS as a negative control.



Please let us know if you have further questions.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (Bone marrow CD34+ cells)
Specification
Bone marrow CD34+ cells
Fixation
Formaldehyde
Permeabilization
Yes - ice-cold methanol drop-wise, then 30min at -20oC, then 0.1% Triton-X in PBS for 15min at room temperature
Gating Strategy
eFlour780 fixative vaibility dye (eBioScience) negative for live cell population
Username

Dr. Bon Ham Yip

Verified customer

Submitted Jul 06 2012

Merci pour votre requête.
Nous n'avons pas mesurée l'affinité de cet anticorps. Je peux par contre vous rassurer que cet anticorps avait été testé en Western blot à une dilution de 1/1000 jusqu'à 1/5000 avec des bons résultats. De plus je peux vou...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (MCF7)
Loading amount
50 µg
Specification
MCF7
Treatment
5-FU, 1mM for 24hrs
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 18°C
Username

Abcam user community

Verified customer

Submitted Mar 07 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (Jurkat)
Specification
Jurkat
Preparation
Cell harvesting/tissue preparation method: Spin down, wash in FACS buffer 1x, fix, wash 2x, and stain with primary antibody overnight.
Sample buffer: Buffer composition at analysis was 0.1% sodium azide with FBS in phosphate buffered solution.
Fixation
Paraformaldehyde
Permeabilization
Yes - Triton X-100 and NP40
Gating Strategy
Isolate cell population from plot of SSC-A / FSA-A
Username

Abcam user community

Verified customer

Submitted Sep 02 2011

1-10 of 11 Abreviews or Q&A

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