SMAC/Diablo Antibody - BSA Free

Western Blot: SMAC/Diablo Antibody [NBP1-76772] - Western Blot Validation in Human, Mouse and Rat Cell Lines. Loading: 15 ug of lysates per lane. Antibodies: SMAC/Diablo NBP1-76772 (1 ug/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Immunohistochemistry-Paraffin: SMAC/Diablo Antibody [NBP1-76772] - Immunohistochemical staining of mouse spleen tissue using Smac antibody at 2 ug/ml.

Western Blot: SMAC/Diablo Antibody [NBP1-76772] - Analysis of Smac in mouse heart tissue lysate in the absence (A) or presence (B) of blocking peptide and in rat heart tissue lysate with Smac antibody at 1 ug /ml.

Western Blot: SMAC/Diablo Antibody [NBP1-76772] - Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines.Loading: 15 ug of lysates per lane. Antibodies: SMAC/Diablo NB100-55900 (1 ug/mL), SMAC/Diablo NBP1-76772 (1 ug/mL), and beta-actin (1 ug/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Immunocytochemistry/ Immunofluorescence: SMAC/Diablo Antibody - BSA Free [NBP1-76772] - Figure 4 Immunofluorescence Validation of SMAC/Diablo in Mouse Spleen CellsImmunofluorescent analysis of 4% paraformaldehyde-fixed Mouse Spleen Cells labeling SMAC/Diablo with at 10 u/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).

Immunohistochemistry: SMAC/Diablo Antibody - BSA Free [NBP1-76772] - Figure 5 Immunohistochemistry Validation of SMAC/Diablo in Mouse Spleen Tissue Immunohistochemical analysis of paraffin-embedded Mouse Spleen Tissue using anti-SMAC/Diablo antibody at 2 u/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, Mu, Rt
• Applications: WB, ELISA, ICC/IF, IHC, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

SMAC/Diablo Antibody - BSA Free Summary

• Immunogen: Antibody was raised against a 16 amino acid peptide near the carboxy terminus of murine Smac. The immunogen is located within the last 50 amino acids of Smac. Amino Acid Squence: SDEGADQEEEAYLRED
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: DIABLO
• Purity: Ion exchange chromatography

Applications/Dilutions

• Dilutions:
  • ELISA 1:100-1:2000
  • Immunocytochemistry/ Immunofluorescence 10 ug/ml
  • Immunohistochemistry 2 ug/mL
  • Immunohistochemistry-Paraffin
  • Immunoprecipitation
  • Western Blot 1 ug/mL
• Theoretical MW: 20 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:

  SMAC/Diablo Peptide (NBP1-76772PEP)

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: Ion exchange chromatography

Alternate Names for SMAC/Diablo Antibody - BSA Free

• 0610041G12Rik
• Diablo
• diablo, IAP-binding mitochondrial protein
• Direct IAP-binding protein with low pI
• mitochondrial Smac protein
• Second mitochondria-derived activator of caspase
• SMAC
• SMACmitochondrial

Background

The inhibitor of apoptosis proteins (IAPs) regulate programmed cell death by inhibiting members of the caspase family of enzymes. A novel mammalian protein that binds to IAPs and neutralizes the inhibitory effect of IAPs on caspases was recently identified and designated Smac/DIABLO. Smac/DIABLO is a mitochondrial protein that is released along with cytochrome c during apoptosis and activates cytochrome c/Apaf-1/caspase-9 pathway. Analysis of the structural basis of Smac/DIABLO reveals that the N-terminal amino acids are required for binding of Smac/DIABLO to IAPs and activation of caspases. Smac/DIABLO is expressed in a variety of human and mouse tissues.

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 SMAC/Diablo Antibody (NBP1-76772)(2)

Publications using NBP1-76772

• Sikorski K, Mehta A et al. A high-throughput pipeline for validation of antibodies. Nat Methods 2018-01-11 [PMID: 30377371] (Human)
  
  Details:
  Antibody validation based on denaturing gel electrophoresis of biotinylated cell lysates (PAGE) followed by mass spectrometry (MS) and antibody array analysis (MAP).
• Sulaiman MK, Chu Z, Blanco VM et al. SapC-DOPS nanovesicles induce Smac- and Bax-dependent apoptosis through mitochondrial activation in neuroblastomas Mol. Cancer 2015-04-19 [PMID: 25889084] (WB, Human)

Bioinformatics

• Gene Symbol: DIABLO
• Uniprot:
  • AF203914(Mouse)