PUMA Antibody - BSA Free

Western Blot: PUMA Antibody - BSA Free [NBP1-76639] - PUMA Antibody [NBP1-76639] - Identification of target for LPS-induced miR-29b in OCs. BMMs were incubated with M-CSF (30 ng/ml) and RANKL (40 ng/ml) for 40 h, washed thoroughly, and incubated further with LPS (50 ng/ml) in the presence of M-CSF (30 ng/ml) for 48 h. Cells were transfected with 30 nM of miR-29b mimic or con mimic in the presence of M-CSF (30 ng/ml) for 6 h. Total RNA was analyzed by qPCR to quantify the expression of BMF, PUMA, BAK1, BIM, and HRK. Expression levels with con mimic treatment were set at 1. Cell lysates were subjected to Western blot analysis with antibodies against BMF, PUMA, BAK1, BIM, HRK, and BCL-2. Antibodies against beta-actin were used for normalization. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/omcl/2019/6018180/) licensed under a CC-BY license.

Western Blot: PUMA Antibody - BSA Free [NBP1-76639] - PUMA Antibody [NBP1-76639] - K562 cell lysate with PUMA antibody at 2 ug/ml.

Western Blot: PUMA Antibody - BSA Free [NBP1-76639] - PUMA Antibody [NBP1-76639] - Independent Antibody Validation (IAV) via Protein Expression ProfileLoading: 20 ug of lysates per lane.Antibodies: NBP1-76654 (3 ug/mL), NBP1-76639 (2 ug/mL), beta-actin (1 ug/mL) and GAPDH (0.02 ug/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate.

Immunocytochemistry/ Immunofluorescence: PUMA Antibody - BSA Free [NBP1-76639] - Validation of PUMA in K562 Cells. Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling PUMA with at 20 ug/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).

Immunocytochemistry/ Immunofluorescence: PUMA Antibody - BSA Free [NBP1-76639] - Validation of PUMA in K562. Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling PUMA with at 10 ug/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red). Image showing cytosol staining on K562 cells

Immunohistochemistry: PUMA Antibody - BSA Free [NBP1-76639] - Validation of PUMA in Human Breast Carcinoma. Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-PUMA antibody at 10 ug/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 4C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunohistochemistry: PUMA Antibody - BSA Free [NBP1-76639] - Validation of PUMA in Human Breast Tissue. Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-PUMA antibody at 2.5 ug/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 4C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

• Reactivity: Hu, Rt
• Applications: WB, ELISA, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

PUMA Antibody - BSA Free Summary

• Immunogen: 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. Amino Acid Squence: ARARQEGSSPEPVEG
• Specificity: 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).
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: BBC3
• Purity: Peptide affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:100-1:2000
  • Immunocytochemistry/ Immunofluorescence 10-20 ug/mL
  • Immunohistochemistry 2.5-10 ug/mL
  • Immunohistochemistry-Paraffin
  • Western Blot 2-3 ug/mL
• Theoretical MW: 23 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control Peptide:

  PUMA Peptide (NBP1-76639PEP)

Reactivity Notes

Rat reactivity reported in scientific literature (PMID: 16399862).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Peptide affinity purified

Alternate Names for PUMA Antibody - BSA Free

• BCL2 binding component 3
• bcl-2-binding component 3
• JFY1
• JFY-1
• p53 up-regulated modulator of apoptosis
• PUMAFLJ42994

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for PUMA Antibody (NBP1-76639)(7)

Publications using NBP1-76639

• Choi SH, Koh DI, Cho SY et al. Temporal and differential regulation of KAISO-controlled transcription by phosphorylated and acetylated p53 highlights a crucial regulatory role of apoptosis J. Biol. Chem. 2019-07-11 [PMID: 31296660] (WB, Human)
• Sul O. J, Rajasekaran M, et al. MicroRNA-29b Enhances Osteoclast Survival by Targeting BCL-2-Modifying Factor after Lipopolysaccharide Stimulation. Oxid Med Cell Longev 2019-05-17 [PMID: 31093317] (WB, Mouse)
• Long JS, Schoonen PM, Graczyk D et al. p73 engages A2B receptor signalling to prime cancer cells to chemotherapy-induced death Oncogene. 2015-02-09 [PMID: 25659586] (WB, Human)
• Fandy TE, Ross DD, Gore SD, Srivastava RK. Flavopiridol synergizes TRAIL cytotoxicity by downregulation of FLIPL. Cancer Chemother Pharmacol. 2007-08-01 [PMID: 17187253] (WB)
  
  Details:
  Puma alpha NT (IMG-458-1). WB: MM1S myeloma cell line treated with flavopiridol, Fig 4A.
• Toth A, Jeffers JR, Nickson P et al. Targeted deletion of Puma attenuates cardiomyocyte death and improves cardiac function during ischemia-reperfusion. Am J Physiol Heart Circ Physiol. 2006-07-01 [PMID: 16399862] (WB, Rat)
  
  Details:
  WB: Fig 1B (primary rat cardiomyocytes exposed to hypoxia, Puma is identified as a 22 kDa band), Fig 2B (primary rat cardiomyoctyes infected with GFP-conjugated Puma-alpha adenoviruses). Note: The specificity of the PUMA IMG-458 antibody was Puma-alpha ad
• Karst AM, Dai DL, Martinka M, Li G. PUMA expression is significantly reduced in human cutaneous melanomas. Oncogene. 2005-02-03 [PMID: 15690057] (IHC-P, Human)
  
  Details:
  Puma (IMG-458). IHC (paraffin): Human dysplastic nevi, primary melanoma, and metastatic melanoma tumor tissues, Fig 1.
• Sarkar S, Biswas SC Studies on Contribution of Astrocyte Secreted Proteins on Neuronal Health in Models of Alzheimer's Disease Thesis 2022-01-01 (WB, Rat)

Bioinformatics

• Gene Symbol: BBC3
• Uniprot:
  • NP_055232(Human)