Smac/DIABLO Polyclonal antibody

• 

  WB analysis using 10434-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10434-1-AP (DIABLO antibody) at dilution of 1:8000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HeLa using 10434-1-AP

  WB result of DIABLO antibody (10434-1-AP; 1:5000; incubated at room temperature for 1.5 hours) with sh-Control and sh-DIABLO transfected HeLa cells.

• 

  IP experiment of HepG2 using 10434-1-AP

  IP result of anti-DIABLO (IP:10434-1-AP, 4ug; Detection:10434-1-AP 1:2000) with HepG2 cells lysate 880 ug.

• 

  IHC staining of mouse testis using 10434-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse testis tissue slide using 10434-1-AP (DIABLO antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of HeLa using 10434-1-AP

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using DIABLO antibody (10434-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  IF Staining of HeLa using 10434-1-AP

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using DIABLO antibody (10434-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

• 

  FC experiment of HeLa using 10434-1-AP

  1X10^6 HeLa cells were intracellularly stained with 0.4 ug Anti-Human DIABLO (10434-1-AP) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Isotype Control. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

• Featured Product
• KD/KO Validated

Smac/DIABLO Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Cat No. 10434-1-AP

Publications(17)

Reviews (1)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (1)

Applications

WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Synonyms

DIABLO, DFNA64, Direct IAP-binding protein with low pI, Diablo IAP-binding mitochondrial protein, cleaved form, Diablo IAP-binding mitochondrial protein

---

Tested Applications

• Positive WB detected in: HEK-293 cells, HeLa cells, HepG2 cells, mouse testis tissue, rat testis tissue
• Positive IP detected in: HepG2 cells
• Positive IHC detected in: mouse testis tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:16000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

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.

Published Applications

• WB: See 15 publications below
• IHC: See 1 publications below
• IF: See 1 publications below

Product Information

10434-1-AP targets Smac/DIABLO in WB, IHC, IF/ICC, FC (Intra), IP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, hamster
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: Smac/DIABLO fusion protein Ag0696
• 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_2092647
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity 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)

Publications

Species	Application	Title
human	WB	Eur J Med Chem; PKM2/PDK1 dual-targeted shikonin derivatives restore the sensitivity of EGFR-mutated NSCLC cells to gefitinib by remodeling glucose metabolism; Authors - Hongyan Lin
mouse	WB	J Cell Physiol; Resveratrol ameliorates podocyte damage in diabetic mice via SIRT1/PGC-1α mediated attenuation of mitochondrial oxidative stress.; Authors - Tao Zhang
	WB	Ann Transl Med; Botrychium schaffneri Underw. extract acts via DIABLO to induce apoptosis and inhibit proliferation of non-small cell lung carcinoma in vitro and in vivo.; Authors - Yu-Jia Liu
human	WB	Chin Med; Shikonin and 4-hydroxytamoxifen synergistically inhibit the proliferation of breast cancer cells through activating apoptosis signaling pathway in vitro and in vivo.; Authors - Hong-Yan Lin
human	WB	Acta Biochim Biophys Sin (Shanghai); Cathelicidin PR-39 peptide inhibits hypoxia/reperfusion-induced kidney cell apoptosis by suppression of the endoplasmic reticulum-stress pathway.; Authors - Jing Liu
rat	IHC	Mol Med Rep; Mechanisms underlying the protective effect of tannic acid against arsenic trioxide‑induced cardiotoxicity in rats: Potential involvement of mitochondrial apoptosis.; Authors - Yucong Xue

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

---

FH

SHUBHAM (Verified Customer) (03-14-2019)

good

• Applications: Western Blot,
• Cell Tissue Type: AC16

Smac/DIABLO Antibody Western Blot, validation ( dilution) in AC16 (Cat no:10434-1-AP)