PUMA Antibody Cat. No.: 3043

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psi-iconSpecifications
HOST SPECIES:Rabbit
SPECIES REACTIVITY: Human, Rat
HOMOLOGY: Predicted species reactivity based on immunogen sequence: Rat: (79%), Mouse: (79%)
IMMUNOGEN: Anti-PUMA antibody (3043) 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.
TESTED APPLICATIONS: ELISA, IF, IHC-P, WB
APPLICATIONS: WB: 2 μg/mL; IF: 10 μg/mL; IHC: 10 μg/mL.

Antibody validated: Western Blot in human samples; Immunohistochemistry in human samples; Immunofluorescence in human and rat samples. All other applications and species not yet tested.
POSITIVE CONTROL:1) Cat. No. 1210 - 293 Cell Lysate
 2) Cat. No. 1211 – HepG2 Cell Lysate
 3) Cat. No. 10-001 - Human Breast Tissue Slide
 4) Cat. No. 1204 – K562 Cell Lysate
PREDICTED MOLECULAR WEIGHT: 23 kDa
psi-iconAdvanced Validation
VALIDATION:

Independent Antibody Validation (Figure 2) shows similar PUMA expression profile in both human and mouse cell lines detected by two independent anti-PUMA antibodies that recognize different epitopes, 3041 against C-terminus and 3043 against the N-terminus.  PUMA proteins are detected in most of the cell lines with different expression levels by the two independent antibodies.  

KO Validation (Figure 7, 8) shows lack of PUMA expression in the thymocytes or neurons of PUMA knockout mice.

Animal Species Reactivity  (Figure 10) shows PUMA expression in the rat retina detected with anti-PUMA antibodies (3043).

ISOFORMS: Human PUMA has 4 isoforms, including isoform 1 (193aa, 21kD), isoform 2 (131aa, 14kD), isoform 3 (101aa, 10kD) and isofrom 4 (261aa, 27kD). This antibody detects human isoform 1, and is predicted to detect mouse and rat PUMA (193aa, 21kD for both of them).

psi-iconProperties
PURIFICATION:PUMA Antibody is affinity chromatography purified via peptide column.
CLONALITY:Polyclonal
ISOTYPE:IgG
CONJUGATE:Unconjugated
PHYSICAL STATE:Liquid
BUFFER:PUMA Antibody is supplied in PBS containing 0.02% sodium azide.
CONCENTRATION:1 mg/mL
STORAGE CONDITIONS:PUMA antibody can be stored at 4˚C for three months and -20˚C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.

psi-iconAdditional Info
OFFICIAL SYMBOL:BBC3
ALTERNATE NAMES:PUMA Antibody: JFY1, PUMA, JFY-1, Bcl-2-binding component 3
ACCESSION NO.:NP_055232
PROTEIN GI NO.:15193488
GENE ID:27113
USER NOTE:Optimal dilutions for each application to be determined by the researcher.
psi-iconBackground and References
BACKGROUND:PUMA Antibody: 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. The PUMA gene encodes two BH3 domain-containing proteins termed PUMA-alpha and PUMA-beta. 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.
REFERENCES:1) Nakano and Vousden. Mol Cell. 2001; 7(3)683-94
2) Han et al. Proc Natl Acad Sci U S A. 2001;98(20):11318-23.
3) Han J, Flemington C, Houghton AB , Gu Z, Zambetti GP, Lutz RJ, Zhu L, Chittenden T. Expression of bbc3, a pro-apoptotic BH3-only gene, is regulated by diverse cell death and survival signals. Proc Natl Acad Sci U S A. 2001;98(20):11318-23.
psi-iconCITATIONS
CITATIONS: 1)Michalak et al. In several cell types tumour suppressor p53 induces apoptosis largely via Puma but Noxa can contribute. Cell Death Differ. 2008;15(6):1019-29. PMID: 18259198
2)Ambacher et al. The JNK- and AKT/GSK3β- signaling pathways converge to regulate Puma induction and neuronal apoptosis induced by trophic factor deprivation. PLoS One. 2012;7(10): e46885. PMID: 23056511
3)Wade et al. BH3 activation blocks Hdmx suppression of apoptosis and cooperates with Nutlin to induce cell death. Cell Cycle. 2008;7(13):1973-82. PMID: 18604177
4)Wakabayashi et al. Prolonged expression of Puma in cholinergic amacrine cells during the development of rat retina. J Histochem Cytochem. 2012;60(10):777-88. PMID: 22736709
5)Charvet et al. Phosphorylation of Tip60 by GSK-3 determines the induction of PUMA and apoptosis by p53. Mol Cell. 2011;42(5):584-96. PMID: 21658600
6)Schubert et al. Requirement of GSK-3 for PUMA induction upon loss of pro-survival PI3K signaling. Cell Death Dis. 2018;9(5):470. PMID: 29686375
7)Jain et al. A phase I-II trial of fludarabine, bendamustine and rituximab (FBR) in previously treated patients with CLL. Oncotarget. 2017;8(13):22104-12. PMID: 27655665
8)Ikonomopoulou et al. Gomesin inhibits melanoma growth by manipulating key signaling cascades that control cell death and proliferation. Sci Rep. 2018 ;8(1):11519. PMID: 30068931

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