Recombinant Anti-Caspase-9 antibody [E84] (ab32068)
![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)")
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, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
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, 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 (glycerin, glycerine), 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
The Abpromise guarantee
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).
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| IHC-P |
1/25. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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| Notes |
|---|
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WB
1/2000. Detects a band of approximately 46 kDa (predicted molecular weight: 46 kDa). |
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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)
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 -
![Anti-Caspase-9 antibody [E84] (ab32068)](https://www.abcam.com/ps/products/32/ab32068/Images/ab32068-5-benefits-of-recombinant-antibodies.png)
Anti-Caspase-9 antibody [E84] (ab32068)
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
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Datasheet download
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
