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rabbit anti-Smac (CT) polyclonal antibody 4798

$445.00

Antibody summary

  • Rabbit polyclonal to Smac (CT)
  • Suitable for: ELISA,WB,IHC-P,IP,IF
  • Isotype: IgG
  • 100 µg
SKU: 4798parent Category: Tags: , ,
Weight1 lbs
Dimensions9 × 5 × 2 in
host

rabbit

isotype

IgG

clonality

polyclonal

concentration

1 mg/mL

applications

ICC/IF, WB

reactivity

Smac (CT)

available sizes

100 µg

Available product – rabbit anti-Smac (CT) polyclonal antibody 4798

antibody
Tested applications
WB,IHC,IHC,ELISA,IP
Recommended dilutions
Immunoblotting: use at 1ug/mL.

Positive control: Human heart tissue lysate.

Immunohistochemistry: use at 5ug/mL

These are recommended concentrations.

Enduser should determine optimal concentrations for their applications.
Immunogen
Synthetic peptide corresponding to aa 225-239 of human Smac (accession no. AAF87716).
Size and concentration
100µg and lot specific
Form
liquid
Storage Instructions
This antibody is stable for at least one (1) year at -20°C. Avoid multiple freeze-thaw cycles.
Storage buffer
PBS, pH 7.4.
Purity
peptide affinty purifcation
Clonality
polyclonal
Isotype
IgG
Compatible secondaries
goat anti-rabbit IgG, H&L chain specific, peroxidase conjugated, conjugated polyclonal antibody 9512
goat anti-rabbit IgG, H&L chain specific, biotin conjugated polyclonal antibody 2079
goat anti-rabbit IgG, H&L chain specific, FITC conjugated polyclonal antibody 7863
goat anti-rabbit IgG, H&L chain specific, Cross Absorbed polyclonal antibody 2371
goat anti-rabbit IgG, H&L chain specific, biotin conjugated polyclonal antibody, crossabsorbed 1715
goat anti-rabbit IgG, H&L chain specific, FITC conjugated polyclonal antibody, crossabsorbed 1720
Isotype control
Rabbit polyclonal - Isotype Control
target relevance
Protein names
Diablo IAP-binding mitochondrial protein (Diablo homolog, mitochondrial) (Direct IAP-binding protein with low pI) (Second mitochondria-derived activator of caspase) (Smac) [Cleaved into: Diablo IAP-binding mitochondrial protein, cleaved form]
Gene names
DIABLO,DIABLO SMAC
Mass
27131Da
Function
FUNCTION: Promotes apoptosis by activating caspases in the cytochrome c/Apaf-1/caspase-9 pathway. Acts by opposing the inhibitory activity of inhibitor of apoptosis proteins (IAP). Inhibits the activity of BIRC6/bruce by inhibiting its binding to caspases. {ECO:0000269|PubMed:10929711, ECO:0000269|PubMed:15200957}.; FUNCTION: [Isoform 3]: Attenuates the stability and apoptosis-inhibiting activity of XIAP/BIRC4 by promoting XIAP/BIRC4 ubiquitination and degradation through the ubiquitin-proteasome pathway. Also disrupts XIAP/BIRC4 interacting with processed caspase-9 and promotes caspase-3 activation. {ECO:0000269|PubMed:14523016}.; FUNCTION: [Isoform 1]: Defective in the capacity to down-regulate the XIAP/BIRC4 abundance. {ECO:0000269|PubMed:14523016}.
Subellular location
SUBCELLULAR LOCATION: Mitochondrion {ECO:0000269|PubMed:10972280, ECO:0000269|PubMed:14523016, ECO:0000269|PubMed:28288130}. Cytoplasm, cytosol {ECO:0000269|PubMed:10929711, ECO:0000269|PubMed:28288130}. Note=Released into the cytosol in a PARL-dependent manner when cells undergo apoptosis. {ECO:0000269|PubMed:10929711, ECO:0000269|PubMed:28288130}.
Tissues
TISSUE SPECIFICITY: Ubiquitously expressed with highest expression in testis. Expression is also high in heart, liver, kidney, spleen, prostate and ovary. Low in brain, lung, thymus and peripheral blood leukocytes. Isoform 3 is ubiquitously expressed. {ECO:0000269|PubMed:10929711, ECO:0000269|PubMed:14523016}.
Structure
SUBUNIT: Homodimer (PubMed:10972280). Interacts with BEX3 (By similarity). Interacts with BIRC2/c-IAP1 (via BIR3 domain) (PubMed:19153467). Interacts with BIRC6/bruce (PubMed:15200957). Interacts with BIRC7/livin (PubMed:16729033). Interacts with XIAP/BIRC4 (via BIR3 domain) (PubMed:21695558, PubMed:11140637, PubMed:28288130). Interacts with the monomeric and dimeric form of BIRC5/survivin (PubMed:21536684). Interacts with AREL1 (via HECT domain); in the cytoplasm following induction of apoptosis (PubMed:23479728). {ECO:0000250|UniProtKB:Q9JIQ3, ECO:0000269|PubMed:10972280, ECO:0000269|PubMed:11140637, ECO:0000269|PubMed:15200957, ECO:0000269|PubMed:16729033, ECO:0000269|PubMed:19153467, ECO:0000269|PubMed:21536684, ECO:0000269|PubMed:21695558, ECO:0000269|PubMed:23479728, ECO:0000269|PubMed:28288130}.
Post-translational modification
PTM: Ubiquitinated by BIRC7/livin. {ECO:0000269|PubMed:16729033}.; PTM: The precursor form is proteolytically cleaved by mitochondrial processing peptidase MPP to remove the transit peptide and produce an intermediate form. This is then processed by PARL to produce the mature cleaved form which is released from mitochondria into the cytosol in apoptotic cells. {ECO:0000269|PubMed:28288130}.
Target Relevance information above includes information from UniProt accession : Q9NR28
The UniProt Consortium

Data

benchmark-antibodies_anti-smac_ct_antibody_4798_1.jpg
Western Blot Validation in Human Heart Tissue Lysate
Loading: 15 µg of lysates per lane. Antibodies: Smac 4798 (1 µg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.(A) Without blocking peptide(B) With blocking peptide
benchmark-antibodies_anti-smac_ct_antibody_4798_2.gif
Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines
Loading: 15 µg of lysates per lane. Antibodies: Smac 4798 (1 µg/mL), Smac 1411 (1 µg/mL), and beta-actin (1 µg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-smac_ct_antibody_4798_3.gif
Western Blot Validation in Human, Mouse and Rat Cell Lines
Loading: 15 µg of lysates per lane. Antibodies: Smac 4798 (1 µg/mL), 1h incubation at RT in 5% NFM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-smac_ct_antibody_4798_4.gif
Western Blot Validation in Mouse 3T3/NIH Cells
Loading: 15 µg of lysate. Antibodies: Smac 4798 (1 µg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-smac_ct_antibody_4798_5.jpg
Immunohistochemistry Validation of Smac in Human Ovary Tissue
Immunohistochemical analysis of paraffin-embedded Human Ovary tissue using anti-Smac antibody (4798) at 5 µg/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.
benchmark-antibodies_anti-smac_ct_antibody_4798_6.gif
KO Validation in Mouse Fibroblasts and Myoblasts (Ho et al., 2416)
The indicated MEFs or MEMs were exposed to 2 uM STS for 4 h and analyzed by Western blot. Accumulation of Smac/Diablo in mitochondrion-depleted cytosol fractions fromSTS-treated Apaf-1 KO cells were detected by anti-smac antibodies. Smac expression was not detected in smac KO mice.
benchmark-antibodies_anti-smac_ct_antibody_4798_7.gif
Immunohistochemistry Validation of Smac in Human gastric carcinoma (Kim et al., 2011)
Smac was highly expressed in gastric mucosa of patients with gastric carcinoma.
benchmark-antibodies_anti-smac_ct_antibody_4798_8.gif
Immunofluorescence Analysis of Smac in NB4-LR1 Cells (Saumet et al., 2005)
NB4-LR1 cells were either treated with ATRA(1 ?M) for 3 days without or with the T3C1 recombinant fragment (3 uM) or treated with staurosporine (STP; 5 uM ) for 3.5 hours. STP, but not ATRA or AYRA/T3C1 induced the release of smac.
benchmark-antibodies_anti-smac_ct_antibody_4798_9.gif
Induced Expression Validation in Rat Liver (Genestier et al., 2005)
Mitochondria from rat liver were treated with increasing concentrations of rPVL (A), rLukS (B), or Bax alpha (C) for 1 hour at 30C. rPVL induces the release of the apoptogenic proteins cytochrome c and Smac/DIABLO from isolated mitochondria.

Publications

Published literature highly relevant to the biological target of this product and referencing this antibody or clone are retrieved from PubMed database provided by The United States National Library of Medicine at the National Institutes of Health.

pmidtitleauthorscitation

Protocols

relevant to this product
Western blot
IHC
ICC

Documents

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