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rabbit anti-Caspase-9 (I120) polyclonal antibody 4094

$445.00

Antibody summary

  • Rabbit polyclonal to Caspase-9 (I120)
  • Suitable for: ELISA,WB,ICC,IF,IP
  • Isotype: IgG
  • 100 µg
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SKU: 4094parent Category: Tags: , ,
Weight1 lbs
Dimensions9 × 5 × 2 in
host

rabbit

isotype

IgG

clonality

polyclonal

concentration

1 mg/mL

applications

ICC/IF, WB

reactivity

Caspase-9 (I20)

available sizes

100 µg

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

Immunocytochemistry: use at 2ug/mL.

These are recommended concentrations.

Enduser should determine optimal concentrations for their applications.

Positive control: Whole cell lysate from HeLa cells.
Immunogen
Peptide corresponding to aa 299- 318 of human caspase-9 (accession no. P55211).
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
Caspase-9 (CASP-9) (EC 3.4.22.62) (Apoptotic protease Mch-6) (Apoptotic protease-activating factor 3) (APAF-3) (ICE-like apoptotic protease 6) (ICE-LAP6) [Cleaved into: Caspase-9 subunit p35; Caspase-9 subunit p10]
Gene names
CASP9,CASP9 MCH6
Protein family
Peptidase C14A family
Mass
46281Da
Function
FUNCTION: Involved in the activation cascade of caspases responsible for apoptosis execution. Binding of caspase-9 to Apaf-1 leads to activation of the protease which then cleaves and activates effector caspases caspase-3 (CASP3) or caspase-7 (CASP7). Promotes DNA damage-induced apoptosis in a ABL1/c-Abl-dependent manner. Proteolytically cleaves poly(ADP-ribose) polymerase (PARP). {ECO:0000269|PubMed:15657060, ECO:0000269|PubMed:16352606, ECO:0000269|PubMed:16916640, ECO:0000269|PubMed:23516580, ECO:0000269|PubMed:27889207, ECO:0000269|PubMed:35338844, ECO:0000269|PubMed:35446120}.; FUNCTION: [Isoform 2]: Lacks activity is an dominant-negative inhibitor of caspase-9. {ECO:0000269|PubMed:10070954}.
Catalytic activity
CATALYTIC ACTIVITY: Reaction=Strict requirement for an Asp residue at position P1 and with a marked preference for His at position P2. It has a preferred cleavage sequence of Leu-Gly-His-Asp-|-Xaa.; EC=3.4.22.62; Evidence={ECO:0000269|PubMed:16916640, ECO:0000269|PubMed:23516580, ECO:0000269|PubMed:27889207};
Tissues
TISSUE SPECIFICITY: Ubiquitous, with highest expression in the heart, moderate expression in liver, skeletal muscle, and pancreas. Low levels in all other tissues. Within the heart, specifically expressed in myocytes. {ECO:0000269|PubMed:16857965}.
Structure
SUBUNIT: Heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 35 kDa (p35) and a 10 kDa (p10) subunit. Caspase-9 and APAF1 bind to each other via their respective NH2-terminal CED-3 homologous domains in the presence of cytochrome C and ATP. Interacts (inactive form) with EFHD2. Interacts with HAX1. Interacts with BIRC2/c-IAP1, XIAP/BIRC4, BIRC5/survivin, BIRC6/bruce and BIRC7/livin. Interacts with ABL1 (via SH3 domain); the interaction is direct and increases in the response of cells to genotoxic stress and ABL1/c-Abl activation. Interacts with BCL2L10 (PubMed:19255499). Interacts with NleF from pathogenic E.coli. {ECO:0000269|PubMed:11734640, ECO:0000269|PubMed:12620238, ECO:0000269|PubMed:15200957, ECO:0000269|PubMed:15657060, ECO:0000269|PubMed:16857965, ECO:0000269|PubMed:19118655, ECO:0000269|PubMed:19255499, ECO:0000269|PubMed:23516580}.
Post-translational modification
PTM: Cleavages at Asp-315 by granzyme B and at Asp-330 by caspase-3 generate the two active subunits. Caspase-8 and -10 can also be involved in these processing events.; PTM: Phosphorylated at Thr-125 by MAPK1/ERK2. Phosphorylation at Thr-125 is sufficient to block caspase-9 processing and subsequent caspase-3 activation. Phosphorylation on Tyr-153 by ABL1/c-Abl; occurs in the response of cells to DNA damage. {ECO:0000269|PubMed:12792650, ECO:0000269|PubMed:15657060}.; PTM: (Microbial infection) ADP-riboxanation by C.violaceum CopC blocks CASP9 processing, preventing CASP9 activation and ability to mediate intrinsic apoptosis. {ECO:0000269|PubMed:35338844, ECO:0000269|PubMed:35446120}.
Target Relevance information above includes information from UniProt accession : P55211
The UniProt Consortium

Data

benchmark-antibodies_anti-caspase-9_i20_antibody_4094_1.gif
Independent Antibody Validation (IAV) via Protein Expression Profile in Human Cell Lines
Loading: 15 µg of lysates per lane. Antibodies: Caspase 9, 2417 (1 µg/mL), Caspase 9, 4094 (1 µg/mL) and beta-actin (1.5 µg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-caspase-9_i20_antibody_4094_2.gif
Western Blot Validation in Human Cell Lines
Loading: 15 µg of lysates per lane. Antibodies: : Caspase 9, 4094 (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-caspase-9_i20_antibody_4094_3.gif
Western Blot Validation in Mouse Cell Line
Loading: 15 µg of 3T3/NIH cell lysate. Antibodies: : Caspase 9, 4094 (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-caspase-9_i20_antibody_4094_4.jpg
Immunofluorescence Validation of Caspase 9 in K562 Cells
Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling Caspase 9 with 4094 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green).
benchmark-antibodies_anti-caspase-9_i20_antibody_4094_5.gif
Immunofluorescence Validation of Caspase 9 in HeLa Cells
Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa cells labeling Caspase 9 with 4094 at 5 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
benchmark-antibodies_anti-caspase-9_i20_antibody_4094_6.gif
Immunocytochemistry Validation of Caspase 9 in HeLa Cells
Immunocytochemical analysis of HeLa cells using anti-Caspase 9 antibody (4094) at 5 µg/mL. Cells 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.

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.

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Protocols

relevant to this product
Western blot
IHC
ICC

Documents

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