Caspase-9 Antibody (LAP6 96-2-22)

Western Blot: Caspase 9 Antibody (LAP6 96-2-22) [NB500-209] - Western blot analysis of Jurkat (A) and HeLa (B) cell lysate using Caspase 9 (NB500-209) antibody at 1:500.

Immunocytochemistry/Immunofluorescence: Caspase 9 Antibody (LAP6 96-2-22) [NB500-209] - The Caspase 9 antibody was tested in HeLa cells against Dylight 488 (Green). Alpha-tubulin and nuclei were counterstained against Dylight 550 (Red) and DAPI (Blue).

Immunohistochemistry-Paraffin: Caspase 9 Antibody (LAP6 96-2-22) [NB500-209] - IHC analysis of paraffin-embedded human breast carcinomausing Caspase 9 antibody at 1:25. Image from verified customer review.

Western Blot: Caspase 9 Antibody (LAP6 96-2-22) [NB500-209] - The pro and active forms of Caspase 9 detected in active 293 cell lysate (lane 2). Lane 1: inactive 293 cell lysate.

Simple Western: Caspase-9 Antibody (LAP6 96-2-22) [NB500-209] - 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 ug/mL of Mouse Anti-Caspase 9 Monoclonal Antibody (Catalog # NB500-209). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

• Reactivity: Hu
• Applications: WB, Simple Western, ICC/IF, IHC, IHC-P, KD
• Clone: LAP6 96-2-22
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

Caspase-9 Antibody (LAP6 96-2-22) Summary

• Immunogen: Recombinant human Caspase 9 pro-domain protein. [UniProt# P55211]
• Localization: Cytoplasmic and Mitochondrial
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: CASP9
• Purity: Protein A or G purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/ml
  • Simple Western 20 ug/ml
  • Immunocytochemistry/Immunofluorescence 2-10 ug/ml
  • Immunohistochemistry 8-25 ug/ml
  • Immunohistochemistry-Paraffin 8-25 ug/ml
  • Knockdown Validated
• Application Notes: This Caspase 9 (LAP6 96-2-22) antibody is useful for Immunocytochemistry/Immunofluorescence and Western Blot, where bands at ~46kDa-48kDa and 35kDa are seen representing the pro and active forms of Caspase 9, respectively. In ICC/IF cytoplasmic staining can be seen in HeLa cells. Paraffin data from customer review.
• Control:

  293 Whole Cell Lysate

Reactivity Notes

Human.

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein A or G purified

Notes

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 ug/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.

Alternate Names for Caspase-9 Antibody (LAP6 96-2-22)

• APAF-3APAF3
• apoptotic protease activating factor 3
• Apoptotic protease Mch-6
• Apoptotic protease-activating factor 3
• CASP9
• CASP-9
• caspase 9, apoptosis-related cysteine peptidase
• Caspase9
• Caspase-9
• EC 3.4.22.62
• ICE-LAP6CASPASE-9c
• ICE-like apoptotic protease 6
• MCH6apoptosis-related cysteine protease

Background

Among the initiator caspases (caspases -2, -8 and -9), caspase-9 is an essential enzymes required for intrinsic or mitochondrial apoptotic pathway. Caspase 9 is a heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 35 kD (p35) and a 10 kD (p10) subunit, and it interacts with several proteins including APAF1, EFHD2, HAX1, BIRC2/c-IAP1, XIAP/BIRC4, BIRC5/survivin, BIRC6/bruce, BIRC7/livin etc. During apoptosome-dependent apoptotic execution, cytochrome-c is released from the mitochondria into cytosol where it binds/activates Apaf-1. Upon nucleotide exchange in the presence of dATP, Apaf-1 oligomerizes into heptameric apoptosome complex and procaspase-9 is recruited to apoptosome where it gets activated. Active caspase-9 at the apoptosome proteolytically activates procaspase-3 (the critical apoptotic executioner caspases) and cleaves poly(ADP-ribose) polymerase (PARP). Both caspases-9 and -3 can be inhibited by selected members of IAPs, with XIAP being their most potent natural inhibitor. Phosphorylation of a major site, Thr125 by CDK1-cyclin B1 leading to inhibition of caspase 9 plays a critical role in the restraint of apoptosis during mitotic arrest. This site is also targeted by ERK1/2 in response to growth factor signalling and by DYRK1A, which plays important roles during development.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Caspase-9 Antibody (NB500-209)(4)

Publications using NB500-209

• Martin, A et al. Apocytochrome c blocks caspase-9 activation and Bax-induced apoptosis. J Biol Chem;277(52):50834-41. 2002 Dec 27. [PMID: 12393884] (WB, Human)
• Wu SH, Chyau CC, Chen JH et al. Anti-cancerous effects of Wasabia japonica extract in Hep3B liver cancer cells via ROS accumulation, DNA damage and p73-mediated apoptosis Journal of Functional Foods 2015 Mar 02 (WB, Human)
• Kim H, Lee KH, Park IA et al. Expression of SIRT1 and apoptosis-related proteins is predictive for lymph node metastasis and disease-free survival in luminal A breast cancer. Virchows Arch. 2015 Aug 18 [PMID: 26280894] (IHC-P, Human)
  
  Details:
  Caspase 9 antibody was used at 1:300 dilution for IHC-P analysis of tissue sections from human cases of luminal A invasive breast ductal carcinoma. The assay was performed on benchmark automatic immunostaining device and the signal detection was performed using biotinylated anti-mouse secondary antibody -peroxidase-labeled streptavidin - DAB method.
• Hsuan SW, Chyau CC, Hung HY et al. The induction of apoptosis and autophagy by Wasabia japonica extract in colon cancer Eur J Nutr 2015 Feb 27 [PMID: 25720497] (WB, Human)

Reviews for Caspase-9 Antibody (NB500-209) (4)

Average Rating: 4.8

(Based on 4 reviews)

We have 4 reviews tested in 1 species: Human.

reviewed by: Anony- mous IHC-P Human 08/29/2014

Summary

• Application: Immunohistochemistry-Paraffin
• Species: Human
• Special Applications: This antibody detected endogenous level of Caspase 9 in human breast carcinoma in IHC at dilution of 1:25.
• File: View PDF

reviewed by: Anony- mous WB Human 08/29/2014

Summary

• Application: Western Blot
• Species: Human
• Special Applications: This antibody detected endogenous level of Caspase 9 in C6 cell lysates in WB at dilution of 1:1000
• File: View PDF

reviewed by: Anonymous WB Human 01/13/2012

Summary

• Application: Western Blot
• Sample Tested: COLO-205 Cell Line
• Species: Human

Blocking

• Blocking Details: Blocking Buffer: 5% non-fat dry milk in TBS

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:500, Primary Ab Incubation Time: overnight

Secondary Antibody

• Secondary Description: Secondary Ab: Goat anti Mouse HRP, Secondary Ab Dilution Ratio: 1:10000

Details

• Detection Notes: Detection Method: ECL, Specific Bands: 35 kDa

reviewed by: Anonymous WB Human 12/23/2011

Summary

• Application: Western Blot
• Sample Tested: Human cell
• Species: Human

Blocking

• Blocking Details: Blocking Buffer: 5% BSA in TBST Buffer

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:1000, Primary Ab Incubation Time: 1 hour

Secondary Antibody

• Secondary Description: Secondary Ab: Goat anti-mouse IgG, HRP conjugated

Details

• Detection Notes: Detection Method: ECL

FAQs for Caspase-9 Antibody (NB500-209). (Showing 1 - 2 of 2 FAQs).

1. Could I use HeLa cell lysate as a positive control for Caspase 9 primary antibodies in WB?
   • After looking through our testing it does appear you should be fine using HeLa as a positive control cell lysate for Caspase 9 in Western Blot, as we have used this sample successfully in our QC tests and shown expression of Caspase 9.
2. Does NB500-209 both recognize human and rat active form of caspase 9 (subunit p35 or subunit p10)? Besides that please let me know if NB500-209 also recognize pro-form caspase 9?
   • Caspase 9 Antibody (LAP6 96-2-22) # NB500-209 is human specific and do not recognize rat samples. It is expected to detect pro as well as activated form of caspase 9. In our QC data using 293 cell lysates, we observed active subunit p35 only not subunit p10. Both of these antibodies were tested on lysates of 293 cells of human origin (also called Human Embryonic Kidney 293 cells). We did not validate them on rat samples as the antibodies are Human specific.

Bioinformatics

• Gene Symbol: CASP9
• Entrez:
  • 842(Human)
• Uniprot:
  • P55211()