Recombinant Anti-Caspase-9 antibody [E84] (ab32068)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E84] to Caspase-9
- Suitable for: WB, Flow Cyt, IHC-P
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-Caspase-9 antibody [E84]
See all Caspase-9 primary antibodies -
Description
Rabbit monoclonal [E84] to Caspase-9 -
Host species
Rabbit -
Tested applications
Suitable for: WB, Flow Cyt, IHC-Pmore details
Unsuitable for: ICC or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Caspase-9 aa 1-100 (N terminal). The exact sequence is proprietary.
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Positive control
- Jurkat cell lysate
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General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
E84 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab32068 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | 1/2000. Detects a band of approximately 46 kDa (predicted molecular weight: 46 kDa). | |
| Flow Cyt | 1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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| IHC-P | 1/25. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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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 caspase-3. Proteolytically cleaves poly(ADP-ribose) polymerase (PARP).
Isoform 2 lacks activity is an dominant-negative inhibitor of caspase-9. -
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. -
Sequence similarities
Belongs to the peptidase C14A family.
Contains 1 CARD domain. -
Developmental stage
Expressed at low levels in fetal heart, at moderate levels in neonate heart, and at high levels in adult heart. -
Post-translational
modificationsCleavages 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. - Information by UniProt
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Database links
- Entrez Gene: 842 Human
- Omim: 602234 Human
- SwissProt: P55211 Human
- Unigene: 329502 Human
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Alternative names
- APAF-3 antibody
- APAF3 antibody
- Apoptosis related cysteine peptidase antibody
see all
Images
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![Western blot - Anti-Caspase-9 antibody [E84] (ab32068)](https://www.abcam.com/ps/products/32/ab32068/Images/ab32068-277443-anti-caspase-9-antibody-e84-western-blot.jpg)
Western blot - Anti-Caspase-9 antibody [E84] (ab32068)Lanes:
Lane 1: Wild-type HAP1 whole cell lysate (40 µg).
Lane 2: CASP9 knockout HAP1 whole cell lysate (40µg).
Lane 3: HeLa whole cell lysate (40 µg).Lanes 1 - 3 Merged signal (red and green). Green - ab32068 observed at 45 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32068 was shown to specifically recognize CASP9 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when CASP9 knockout samples were examined. Wild-type and CASP9 knockout samples were subjected to SDS-PAGE. Ab32068 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-Caspase-9 antibody [E84] (ab32068)](https://www.abcam.com/ps/products/32/ab32068/Images/ab32068-279861-anti-caspase-9-antibody-e84-western-blot.png)
Western blot - Anti-Caspase-9 antibody [E84] (ab32068)All lanes : Anti-Caspase-9 antibody [E84] (ab32068) at 1/1000 dilution
Lane 1 : Untreated HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa treated with staurosporine 1uM for 4 hours whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 46 kDa
Additional bands at: 35 kDa, 37 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 3 minutesBlocking/dilution buffer: 5% NFDM/TBST.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E84] (ab32068)](https://www.abcam.com/ps/products/32/ab32068/Images/ab32068-43789-anti-caspase-9-antibody-e84-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E84] (ab32068)Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using anti-Pro-Caspase-9 RabMAb (ab32068).
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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![Western blot - Anti-Caspase-9 antibody [E84] (ab32068)](https://www.abcam.com/ps/products/32/ab32068/Images/ab32068-43790-anti-caspase-9-antibody-e84-western-blot.jpg)
Western blot - Anti-Caspase-9 antibody [E84] (ab32068)Anti-Caspase-9 antibody [E84] (ab32068) at 1/2000 dilution + Jurkat cell lysate at 10 µg
Predicted band size: 46 kDa
Observed band size: 46 kDa
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
References (11)
ab32068 has been referenced in 11 publications.
- Yang Q et al. Lidocaine alleviates cytotoxicity-resistance in lung cancer A549/DDP cells via down-regulation of miR-21. Mol Cell Biochem 456:63-72 (2019). PubMed: 30644017
- Zhao J et al. Salidroside mitigates hydrogen peroxide-induced injury by enhancement of microRNA-27a in human trabecular meshwork cells. Artif Cells Nanomed Biotechnol 47:1758-1765 (2019). PubMed: 31062616
- Sun T et al. Knockdown of Long Non-Coding RNA RP11-445H22.4 Alleviates LPS-Induced Injuries by Regulation of MiR-301a in Osteoarthritis. Cell Physiol Biochem 45:832-843 (2018). PubMed: 29414810
- Gu LL et al. Oxymatrine Causes Hepatotoxicity by Promoting the Phosphorylation of JNK and Induction of Endoplasmic Reticulum Stress Mediated by ROS in LO2 Cells. Mol Cells 41:401-412 (2018). PubMed: 29754474
- Liu Y & Zhang Y Lycium barbarum polysaccharides alleviate hydrogen peroxide-induced injury by up-regulation of miR-4295 in human trabecular meshwork cells. Exp Mol Pathol 106:109-115 (2018). PubMed: 30594603
