Smac/DIABLO Monoclonal antibody

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  WB analysis using 68480-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 68480-1-Ig (DIABLO antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control.

• 

  WB analysis using 68480-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 68480-1-Ig (DIABLO antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IF Staining of HepG2 using 68480-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using Smac/DIABLO antibody (68480-1-Ig, Clone: 2F12C9 ) at dilution of 1:800 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) (SA00013-1), CL594-phalloidin (red).

Smac/DIABLO Monoclonal Antibody for WB, IF/ICC, ELISA

Cat No. 68480-1-Ig

Clone No.2F12C9

Host / Isotype

Mouse / IgG1

Reactivity

human, rat

Applications

WB, IF/ICC, ELISA

Synonyms

DIABLO, 2F12C9, DFNA64, Diablo homolog, mitochondrial, Diablo IAP-binding mitochondrial protein

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Tested Applications

• Positive WB detected in: U2OS cells, K-562 cells, LNCaP cells, HeLa cells, HEK-293 cells, Jurkat cells, HSC-T6 cells
• Positive IF/ICC detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:400-1:1600

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

68480-1-Ig targets Smac/DIABLO in WB, IF/ICC, ELISA applications and shows reactivity with human, rat samples.

• Tested Reactivity: human, rat
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag32774

  Product name: Recombinant human DIABLO protein

  Source: e coli.-derived, PET28a

  Tag: 6*His

  Domain: 1-239 aa of BC011909

  Sequence: MAALKSWLSRSVTSFFRYRQCLCVPVVANFKKRCFSELIRPWHKTVTIGFGVTLCAVPIAQKSEPHSLSSEALMRRAVSLVTDSTSTFLSQTTYALIEAITEYTKAVYTLTSLYRQYTSLLGKMNSEEEDEVWQVIIGARAEMTSKHQEYLKLETTWMTAVGLSEMAAEAAYQTGADQASITARNHIQLVKLQVEEVHQLSRKAETKLAEAQIEELRQKTQEEGEERAESEQEAYLRED

• Full Name: diablo homolog (Drosophila)
• Calculated Molecular Weight: 27 kDa
• Observed Molecular Weight: 20 kDa
• GenBank Accession Number: BC011909
• Gene Symbol: DIABLO
• Gene ID (NCBI): 56616
• RRID: AB_3085191
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• UNIPROT ID: Q9NR28
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Smac/DIABLO is a mitochondrial protein that promotes apoptosis by neutralizing members of the IAP family. SMAC/DIABLO is ubiquitously expressed in normal tissues, and the expression of SMAC messenger RNA (mRNA) in adult testis is the highest .Smac/DIABLO encodes a 239 amino acid protein (~27 kDa). The first 55 amino acids are a mitochondrial targeting signal peptide that is cleaved to generate mature Smac-DIABLO (~21 kDa). When released from mitochondria, Smac-DIABLO acts as a natural antagonist of inhibitor of apoptosis proteins (IAPs). Smac-DIABLO antagonistic activity is based on its N-terminal tetrapeptide (AVPI) that binds baculoviral IAP repeat (BIR) domains of IAPs, releasing their inhibitory effects on both initiator and effector caspases, thus promoting cell death. (PMID: 32325691, PMID: 32575872,PMID: 25650938)