Invitrogen

Caspase 9 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

3 Published Figures

3 References

View all (60) Caspase 9 antibodies

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Protocols

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Caspase 9 Antibody in Immunocytochemistry (ICC/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 Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry (Paraffin) analysis of Caspase 9 was performed in paraffin-embedded human lung cancer tissue using Caspase 9 Polyclonal Antibody (Product # PA5-22252) at a dilution of 1:400.

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 & 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 — 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

PA5-22252

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:3,000	View 3 publications 3 publications
Immunohistochemistry (Paraffin) (IHC (P))	1:100-1:1,000	-
Immunocytochemistry (ICC/IF)	5 µg/mL	-
Immunoprecipitation (IP)	Assay-dependent	-

Product Specifications
Species Reactivity	Human, Rat
Published species	Human
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) View antigen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.55 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS, pH 7, with 20% glycerol
Contains	0.025% Proclin 300
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

Recommended positive controls: HeLa (30 µM cisplatin treatment for 24 hr), NTUB1.

Predicted reactivity: Dog (80%), Bovine (82%).

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

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).

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

Product References

Anti-Apoptotic Effect of Apelin in Human Placenta: Studies on BeWo Cells and Villous Explants from Third-Trimester Human Pregnancy.

International journal of molecular sciences

Mlyczyńska E,Myszka M,Kurowska P,Dawid M,Milewicz T,Bałajewicz-Nowak M,Kowalczyk P,Rak A

Published figure using Caspase 9 polyclonal antibody (Product # PA5-22252) in Western Blot

Tue Mar 09 00:00:00 EST 2021

FAT4 silencing promotes epithelial-to-mesenchymal transition and invasion via regulation of YAP and β-catenin activity in ovarian cancer.

BMC cancer

Malgundkar SH,Burney I,Al Moundhri M,Al Kalbani M,Lakhtakia R,Okamoto A,Tamimi Y

PA5-22252 was used in Western Blotting to examine the role of FAT4 in proliferation and metastasis, and its mechanisms of interaction in these processes.

Mon May 04 00:00:00 EDT 2020

FAT4 silencing promotes epithelial-to-mesenchymal transition and invasion via regulation of YAP and β-catenin activity in ovarian cancer.

BMC cancer

Malgundkar SH,Burney I,Al Moundhri M,Al Kalbani M,Lakhtakia R,Okamoto A,Tamimi Y

PA5-22252 was used in Western Blotting to examine the role of FAT4 in proliferation and metastasis, and its mechanisms of interaction in these processes.

Mon May 04 00:00:00 EDT 2020

In Vitro Effects of Vaspin on Porcine Granulosa Cell Proliferation, Cell Cycle Progression, and Apoptosis by Activation of GRP78 Receptor and Several Kinase Signaling Pathways Including MAP3/1, AKT, and STAT3.

International journal of molecular sciences

Kurowska P,Mlyczyńska E,Dawid M,Opydo-Chanek M,Dupont J,Rak A

PA5-22252 was used in Western Blotting to examine the effect of vaspin on granulosa (Gc) proliferation, cell cycle regulation, and apoptosis.

Tue Nov 19 00:00:00 EST 2019

Bioinformatics

Protein Aliases: 25 kDa caspase-9 dominant negative protein; APAF-3; apoptotic protease activating factor 3; Apoptotic protease Mch-6; Apoptotic protease-activating factor 3; cas9; CASP 9; CASP-9; caspase 9, apoptosis-related cysteine peptidase; Caspase-9; caspase-9 dominant negative form; Caspase-9 precursor; Caspase-9 subunit p10; Caspase-9 subunit p35; caspase-9-carboxyl-terminal divergent; CASPASE-9c; Caspase9; CTD; ICE-LAP6; ICE-like apoptotic protease 6; OTTHUMP00000002324; protein phosphatase 1, regulatory subunit 56; RNCASP9; RP11-265F14.3

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

UniProt ID: (Human) P55211

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

Molecular Function: protease