Invitrogen

Caspase 9 Polyclonal Antibody

Advanced Verification

This Antibody was verified by KnockDown to ensure that the antibody binds to the antigen stated. View Details

View all (23) Caspase 9 antibodies

Datasheet

Datasheet

Product Image Gallery

Caspase 9 Antibody in Immunofluorescence (IF) — Immunofluorescence analysis of Caspase 9 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Caspase 9 Rabbit Polyclonal Antibody(Product # PA5-22252) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Nucleus and cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Caspase 9 Antibody in Western Blot (WB) — Knockdown of Caspase 9 was achieved by transfecting HeLa cells with Caspase 9 specific siRNAs (Silencer® select Product # s2429 and s2430). Western blot analysis (Fig. a) was performed using membrane enriched extracts from the Caspase 9 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-Caspase 9 Polyclonal Antibody (Product # PA5-22252, 1:500 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Caspase 9.

Caspase 9 Antibody in KnockDown — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HeLa cells were transfected with Caspase 9 siRNA and loss of signal was observed in Western Blot using Anti-Caspase 9 Polyclonal Antibody (Product # PA5-22252). {KD}

Product Details

Tested Applications

Dilution

Immunocytochemistry (ICC)

5 µg/mL

Immunofluorescence (IF)

5 µg/mL

Western Blot (WB)

1:500

Product Specifications

Species Reactivity

Human, Rat

Host / Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Recombinant fragment corresponding to a region within amino acids 88 and 416 of Caspase 9 (Uniprot ID#P55211)

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Antigen affinity chromatography

Storage buffer

0.1M tris glycine, pH 7, with 10% glycerol

Contains

0.01% thimerosal

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_11152725

Product Specific Information

PA5-22252 targets Caspase 9 in WB applications and shows reactivity with Human samples.

The PA5-22252 immunogen is a recombinant fragment corresponding to a region within amino acids 88 and 416 of Caspase 9 (Uniprot ID#P55211).

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial. For short-term storage (1-2 weeks), product can be stored at 4°C. For long-term storage, aliquot and store product at -20° C or below, avioiding multiple freeze-thaw cycles.

Target Information

Caspase 9 (ICE-like apoptotic protease 6, ICE LAP6, apoptotic protease Mch6, apoptotic protease activating factor 3, Apaf3) is a member of the peptidase family C14 that contains a CARD domain. Caspases are cysteine proteases, expressed as inactive precursors, that mediate apoptosis by proteolysis of specific substrates. Caspase 9 is active as a heterotetramer, is present in the cytosol and, upon activation, translocates to the mitochondria. Caspase 9 is involved in the caspase activation cascade responsible for apoptosis execution and cleaves/activates Caspase 3 and Caspase 6. Further, Caspase 9 is 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 caspase-3. Caspase 9 promotes DNA damage-induced apoptosis in a ABL1/c-Abl-dependent manner, and proteolytically cleaves poly(ADP- ribose) polymerase (PARP). Caspase 9 is expressed in variety of human tissues. An isoform of rat Caspase-9 has been identified in which the C terminus of full-length Caspase-9 is replaced with an alternative peptide sequence. Mutations in Caspase 9 affect brain development. In mice, insufficient Caspase 9 have resulted in abnormal cerebrum development and neuron development. In humans, dysfunctional Caspase 9 expression vary from tissue to tissue. Low levels of Caspase 9 may play a role in cancer development and neurodegenerative diseases such as Alzheimer's Disease (AD).

⚠WARNING: This product can expose you to chemicals including mercury, which is known to the State of California to cause birth defects or other reproductive harm. For more information go to www.P65Warnings.ca.gov.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: 25 kDa caspase-9 dominant negative protein; APAF-3; APAF3; apoptotic protease activating factor 3; Apoptotic protease Mch-6; Apoptotic protease-activating factor 3; cas9; CASP-9; caspase 9, apoptosis-related cysteine peptidase; Caspase-9; caspase-9 dominant negative form; Caspase-9 subunit p10; Caspase-9 subunit p35; caspase-9-carboxyl-terminal divergent; CASPASE-9c; ICE-LAP6; ICE-like apoptotic protease 6; MCH6; PPP1R56; protein phosphatase 1; protein phosphatase 1, regulatory subunit 56; regulatory subunit 56; RP11-265F14.3

Gene Aliases: APAF-3; APAF3; Casp-9-CTD; CASP9; Casp9_v1; ICE-LAP6; MCH6; PPP1R56

UniProt ID: (Human) B4E1A3

Entrez Gene ID: (Human) 842, (Rat) 58918

Molecular Function: cysteine protease enzyme modulator hydrolase protease protease inhibitor