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Purified anti-BBC3/PUMA Antibody

Pricing & Availability
Clone
W22198A (See other available formats)
Regulatory Status
RUO
Other Names
Bcl-2-binding component 3, isoforms 1/2, BCL2 binding component 3 , JFY-1 , p53 up-regulated modulator of apoptosis
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
a. W22198A_PURE_PUMA_WB_062425
Whole cell extracts (15 µg total protein per lane) from the indicated cell lines were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane, and probed with Purified anti-BBC3/PUMA (clone W22198A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-β-actin (Cat. No. 664804) was used as a loading control at a 1:5000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • a. W22198A_PURE_PUMA_WB_062425
    Whole cell extracts (15 µg total protein per lane) from the indicated cell lines were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane, and probed with Purified anti-BBC3/PUMA (clone W22198A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-β-actin (Cat. No. 664804) was used as a loading control at a 1:5000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • b. W22198A_PURE_PUMA_ICC_062425
    HepG2 cells were either untreated (mock) or treated with 500 nM Doxorubicin for 24H (DOX) to induce apoptosis and BBC3 expression. The mock and treated HepG2 cells along with PC-3 cells (negative for BBC3) were fixed with ice-cold methanol and then blocked with 5% FBS for 1 hour at room temperature. Cells were then intracellularly stained with Purified Rat IgG2b, κ Isotype Control Antibody (Cat. No. 400602) (panels A, D, G) or stained with Purified anti-BBC3/PUMA (clone W22198A) (panels B, E, H). Secondary staining was done by incubation with Alexa Fluor® 488 Goat anti-rat IgG (Cat. No. 405418) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801) (blue, merged in panels C, F, I). The image was captured with a 40X objective. Scale bar: 25 μm
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649451 25 µg $165
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649452 100 µg $465
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Description

BBC3, also known as p53 upregulated modulator of apoptosis (PUMA), is a member of the Bcl-2 family. BBC3 promotes apoptosis and can be transcriptionally regulated by p53. When p53 is activated by survival signals, the increased levels of BBC3 induce apoptosis through mitchondrian dysfunction where BBC3 proteins are localized. BBC3 can also induce apoptosis in a p53-independent manner due to other stimuli such as cell stress. p53-mediated apoptosis and tumor suppression are impaired by targeted disruption of the BBC3 gene. There are four different isoforms of BBC3 (α, β, γ, and δ) but only the α and β forms interact with Bcl-2 and Bcl-xL, localize to the mitochondria, and promote apoptosis.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Recombinant human BBC3 protein
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.125 - 1.0 µg/mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This antibody (clone W22198A) does not cross-react with mouse BBC3. This product is not suggested for use in mouse.

Additional Product Notes

For immunocytochemistry (ICC), we recommend to use 4% PFA fixation followed by permeabilization with ice cold-methanol, or ice-cold methanol fixation.

This antibody (clone W22198A) is not suitable for IHC.

RRID
AB_3717147 (BioLegend Cat. No. 649451)
AB_3717147 (BioLegend Cat. No. 649452)

Antigen Details

Structure
BBC3 is a 193 amino acid protein with a predicted molecular weight of 21 kD.
Distribution

Mitochondria

Function
Apoptosis
Biology Area
Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Cell Death
Antigen References
  1. Yu J, et al. 2001. Mol Cell. 7:673.
  2. Nakano K and Vousden K.H. 2001. Mol Cell. 7:683.
  3. Bouillet P and Strasser A. 2002. J Cell Sci. 115:1567.
  4. Jeffers JR, et al. 2003. Cancer Cell. 4:321.
  5. Hemann M.T, et al. 2004. Proc Natl Acad Sci USA. 101:9333.
  6. Yu J, et al. 2003. Proc Natl Acad Sci USA. 100:1931.
  7. Villunger A, et al. 2003. Science. 302:1036.
  8. Roufayel R, et al. 2022. Life (Basel). 12:256.
Gene ID
27113 View all products for this Gene ID
UniProt
View information about BBC3 on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 1    Revision Date: 06/24/2025

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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