Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunohistochemistry, Western Blot, Simple Western

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # LAP6
View all Caspase-9 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human Caspase-9
aa 1-134

Specificity

Detects human Caspase-9 in Western blots and captures Caspase-9 complexed with APAF-1.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human Caspase‑9 Antibody

Capture of Human Caspase-9 and Human Caspase-9 complexed with APAF-1 detected by Western Blot.

Capture of Human Caspase-9 and Human Caspase-9 complexed with APAF-1 detected by Western Blot.

Western blot shows Jurkat human acute T cell leukemia cell line lysates untreated (-) or treated (+) with 50 mM dATP and 1 mg/mL rat cytochrome c for 60 minutes, then captured on a 6-well dish coated at 10 µg/mL with Mouse Anti-Human Caspase-9 Monoclonal Antibody (Catalog # MAB8301). PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Caspase-9 Monoclonal Antibody (Catalog # MAB8301, left side) or Mouse Anti-Human APAF-1 Monoclonal Antibody (MAB868, right side) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF007). Specific bands were detected for Caspase-9 Precursor at approximately 46 kDa and the Caspase-9 p37 subunit at approximately 37 kDa (as indicated). A specific band was detected for APAF-1, captured as part of Caspase-9 complexed with APAF-1, at approximately 135 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.

Detection of Human Caspase-9 antibody by Simple WesternTM.

Detection of Human Caspase‑9 by Simple WesternTM.

Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 mM Staurosporine (STS) for 3 hours, loaded at 0.2 mg/mL. A specific band was detected for Caspase‑9 at approximately 53 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human Caspase‑9 Monoclonal Antibody (Catalog # MAB8301). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

Detection of Human Caspase‑9 by Western Blot.

Western blot shows lysates of HEK293T human embryonic kidney cell line and HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Caspase‑9 Monoclonal Antibody (Catalog # MAB8301) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Caspase‑9 at approximately 46 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 4.

Detection of Human Caspase‑9 by Western Blot.

Western blot shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 ug/ml Staurosporine (STS) for 2 hours. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Caspase‑9 Monoclonal Antibody (Catalog # MAB8301) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Caspase‑9 at approximately 46, 37, 35 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 4.

Detection of Human Caspase-9 by Western Blot

Detection of Human Caspase-9 by Western Blot

Sorbitol-dependent TDP-43 relocalization is reversible and not dependent on Nup153 cleavage.(A) Western blot of nuclear pore complex protein Nup153, nuclear lamina component lamin B1, procaspases -3 and -9 and their activated forms caspase-3 and caspase-9 in HeLa cells treated with sorbitol after pre-treatment with DMSO control or caspase inhibitor Z-VAD-FMK. alpha -tubulin was used as a loading control. (B) Immunofluorescence staining of nuclear-cytoplasmic shuttling proteins TDP-43 (green) and HuR (red) in sorbitol-stressed HeLa cells after 0 min, 15 min or 60 min rescue in normal medium. Scale bar = 30 μm. (C) Quantification of cytoplasmic TDP-43 and HuR protein in sorbitol-treated HeLa cells after 0 min, 15 min or 60 min rescue (N = 3). Results represent mean percentage of cytoplasmic signal ± SEM;** p < 0.01, *** p < 0.001, **** p < 0.0001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28510586), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Human Caspase-9 Antibody by Western Blot

Detection of Human Human Caspase-9 Antibody by Western Blot

Sorbitol-dependent TDP-43 relocalization is reversible and not dependent on Nup153 cleavage.(A) Western blot of nuclear pore complex protein Nup153, nuclear lamina component lamin B1, procaspases -3 and -9 and their activated forms caspase-3 and caspase-9 in HeLa cells treated with sorbitol after pre-treatment with DMSO control or caspase inhibitor Z-VAD-FMK. alpha -tubulin was used as a loading control. (B) Immunofluorescence staining of nuclear-cytoplasmic shuttling proteins TDP-43 (green) and HuR (red) in sorbitol-stressed HeLa cells after 0 min, 15 min or 60 min rescue in normal medium. Scale bar = 30 μm. (C) Quantification of cytoplasmic TDP-43 and HuR protein in sorbitol-treated HeLa cells after 0 min, 15 min or 60 min rescue (N = 3). Results represent mean percentage of cytoplasmic signal ± SEM;** p < 0.01, *** p < 0.001, **** p < 0.0001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28510586), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Caspase-9 by Western Blot

Detection of Human Caspase-9 by Western Blot

Sorbitol-dependent TDP-43 relocalization is reversible and not dependent on Nup153 cleavage.(A) Western blot of nuclear pore complex protein Nup153, nuclear lamina component lamin B1, procaspases -3 and -9 and their activated forms caspase-3 and caspase-9 in HeLa cells treated with sorbitol after pre-treatment with DMSO control or caspase inhibitor Z-VAD-FMK. alpha -tubulin was used as a loading control. (B) Immunofluorescence staining of nuclear-cytoplasmic shuttling proteins TDP-43 (green) and HuR (red) in sorbitol-stressed HeLa cells after 0 min, 15 min or 60 min rescue in normal medium. Scale bar = 30 μm. (C) Quantification of cytoplasmic TDP-43 and HuR protein in sorbitol-treated HeLa cells after 0 min, 15 min or 60 min rescue (N = 3). Results represent mean percentage of cytoplasmic signal ± SEM;** p < 0.01, *** p < 0.001, **** p < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28510586), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Caspase-9 by Western Blot

Detection of Human Caspase-9 by Western Blot

Sorbitol-dependent TDP-43 relocalization is reversible and not dependent on Nup153 cleavage.(A) Western blot of nuclear pore complex protein Nup153, nuclear lamina component lamin B1, procaspases -3 and -9 and their activated forms caspase-3 and caspase-9 in HeLa cells treated with sorbitol after pre-treatment with DMSO control or caspase inhibitor Z-VAD-FMK. alpha -tubulin was used as a loading control. (B) Immunofluorescence staining of nuclear-cytoplasmic shuttling proteins TDP-43 (green) and HuR (red) in sorbitol-stressed HeLa cells after 0 min, 15 min or 60 min rescue in normal medium. Scale bar = 30 μm. (C) Quantification of cytoplasmic TDP-43 and HuR protein in sorbitol-treated HeLa cells after 0 min, 15 min or 60 min rescue (N = 3). Results represent mean percentage of cytoplasmic signal ± SEM;** p < 0.01, *** p < 0.001, **** p < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28510586), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Caspase-9 by Western Blot

Detection of Human Caspase-9 by Western Blot

Sorbitol-dependent TDP-43 relocalization is reversible and not dependent on Nup153 cleavage.(A) Western blot of nuclear pore complex protein Nup153, nuclear lamina component lamin B1, procaspases -3 and -9 and their activated forms caspase-3 and caspase-9 in HeLa cells treated with sorbitol after pre-treatment with DMSO control or caspase inhibitor Z-VAD-FMK. alpha -tubulin was used as a loading control. (B) Immunofluorescence staining of nuclear-cytoplasmic shuttling proteins TDP-43 (green) and HuR (red) in sorbitol-stressed HeLa cells after 0 min, 15 min or 60 min rescue in normal medium. Scale bar = 30 μm. (C) Quantification of cytoplasmic TDP-43 and HuR protein in sorbitol-treated HeLa cells after 0 min, 15 min or 60 min rescue (N = 3). Results represent mean percentage of cytoplasmic signal ± SEM;** p < 0.01, *** p < 0.001, **** p < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28510586), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Caspase‑9 Antibody

Application
Recommended Usage

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human colon

Simple Western

20 µg/mL
Sample: Jurkat human acute T cell leukemia cell line treated with Staurosporine (STS)

Western Blot

1 µg/mL
Sample: Jurkat cells treated with staurosporine (STS), Jurkat cells treated with dATP and rat cytochrome c, HEK293T cells, HepG2 cells

Reviewed Applications

Read 1 review rated 5 using MAB8301 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

=
÷

Background: Caspase-9

Caspase-9 (Cysteine-aspartic acid protease 9/Casp-9; also APAF-3, Mch6 and ICE-LAP6) is a 35-37 kDa member of the peptidase C14A family of enzymes. Casp-9 is an initiator caspase that is part of the intrinsic apoptosis pathway. It is widely expressed and is particularly important during development. Human proCaspase-9 is a 47-48 kDa, 416 amino acid (aa) protein and it contains one CARD region (aa 1-92) and catalytic residues at His237 and Cys287. Following mitochondrial disruption, cytochrome c is released from mitochrondria. Cytochrome c acts on APAF-1, which induces procaspase-9 dimerization. The act of dimerization activates proCasp-9, leading to either the activation of Casp-3, or the autocleavage of proCasp-9, generating a 35 kDa subunit (aa 1-315) and a 12 kDa subunit. Activated Casp-3 will also act on proCasp-9, generating a 37 kDa subunit (aa 1-330) and a 10 kDa subunit (aa 331-416). These subunits associate to form an active heterotetramer. Casp-9 has an alternative start site at Met84 and a deletion of aa 140-289 that generates a dominant negative, 31 kDa isoform. Over aa 1-134, human Casp-9 shares 81% aa identity with mouse Casp-9.

Alternate Names

CASP9, Caspase9

Entrez Gene IDs

842 (Human)

Gene Symbol

CASP9

Additional Caspase-9 Products

Product Documents for Human Caspase‑9 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Human Caspase‑9 Antibody

For research use only

Related Research Areas

Cysteine Proteases and Regulators

Citations for Human Caspase‑9 Antibody

Customer Reviews for Human Caspase‑9 Antibody (1)

5 out of 5
1 Customer Rating
5 Stars
100%
4 Stars
0%
3 Stars
0%
2 Stars
0%
1 Stars
0%

Have you used Human Caspase‑9 Antibody?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Customer Images

Showing  1 - 1 of 1 review Showing All
Filter By:
Clear Filters X
  • Human Caspase-9 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: MDA-MB-231 human breast cancer cell line
    Species: Human
    Verified Customer | Posted 01/17/2018
    Human Caspase‑9 Antibody MAB8301

There are no reviews that match your criteria.

Showing  1 - 1 of 1 review Showing All

Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs

No product specific FAQs exist for this product.

View all FAQs for Antibodies