Anti-PUMA BBC3 Antibody

Product Name

Anti-PUMA BBC3 Antibody

View all PUMA Antibodies

SKU/Catalog Number

A04899-1

Size

0.1 mg

Form

Liquid

Description

Boster Bio Anti-PUMA BBC3 Antibody (Catalog # A04899-1). Tested in ELISA, WB, IHC-P, IF applications. This antibody reacts with Human, Rat.

Storage & Handling

PUMA antibody can be stored at 4°C for three months and -20°C, stable for up to one year. Avoid repeated freeze-thaw cycles. Antibodies should not be exposed to prolonged high temperatures.

Cite This Product

Anti-PUMA BBC3 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # A04899-1)

Host

Rabbit

Contents

PUMA Antibody is supplied in PBS containing 0.02% sodium azide.

Clonality

Polyclonal

Isotype

IgG

Immunogen

Anti-PUMA antibody was raised against a peptide corresponding to 14 amino acids near the amino terminus of human PUMA isoform 1. The immunogen is located within the first 50 amino acids of PUMA.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

A lower band at approximately 16 kDa was detected in Daudi and K562 cells, which may represent the PUMA isoform 2.

Reactive Species

A04899-1 is reactive to BBC3 in Human, Rat

Applications

A04899-1 is guaranteed for ELISA, IF, IHC-P, WB Boster Guarantee

Observed Molecular Weight

68 kDa

Calculated molecular weight

26.498kDa

Background of PUMA

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. PUMA/bbc3 is one of the pro-apoptotic Bcl-2 family members including Bax and Noxa, which are also transcriptional targets of p53 (1). The PUMA gene encodes two BH3 domain-containing proteins termed PUMA-alpha and PUMA-beta (2). 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.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

WB: 2-3 μg/mL; IF: 10-20 μg/mL; IHC: 2.5-10 μg/mL.
Antibody validated: Western Blot in human samples; Immunohistochemistry in human samples; Immunofluorescence in human samples. All other applications and species not yet tested. Optimal dilutions for each application should be determined by the researcher.

Validation Images & Assay Conditions

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Western Blot Validation of PUMA in K562 Cells
Loading: 15 μg of lysates per lane.Antibodies: A04899-1 (2 μg/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution

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Independent Antibody Validation (IAV) via Protein Expression Profile in Human Cells
Loading: 20 μg of lysates per lane.Antibodies: (3 μg/mL), A04899-1 (2 μg/mL), beta-actin (1 μg/mL) and GAPDH (0.02 μg/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

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Immunofluorescence Validation of PUMA in K562 Cells
Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling PUMA with A04899-1 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).

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Immunohistochemistry Validation of PUMA in Human Breast Carcinoma
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-PUMA antibody (A04899-1) at 10 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

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Immunohistochemistry Validation of PUMA in Human Breast Tissue
Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-PUMA antibody (A04899-1) at 2.5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

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Immunofluorescence Validation of PUMA in K562
Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling PUMA with A04899-1 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red). Image showing cytosol staining on K562 cells

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KO Validation of PUMA in Mouse Thymocytes (Michalak et al.

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KO Validation of PUMA in Mouse Cerebellar Neurons (Ambacher et al.

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Induced Expression of PUMA in MCF7 cells (Wade et al.

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Immunofluorescence Validation of PUMA in Rat Retina (Wakabayashi et al., 2012)
PUMA expression in the rat retina detected by anti-PUMA antibodies (A04899-1). The specimens were counterstained with Hoechst 33258 to visualize nuclei (+DNA). GCL, ganglion cell layer; INL, inner nuclear layer; IPL, inner plexiform layer; ONL, outer nuclear layer; OPL, outer plexiform layer; P, postnatal day.