Anti-Caspase-9 antibody [E84] (ab32068)
- Datasheet
- References (8)
- Protocols
Overview
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Product nameAnti-Caspase-9 antibody [E84]
See all Caspase-9 primary antibodies -
DescriptionRabbit monoclonal [E84] to Caspase-9
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Host speciesRabbit
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Tested applicationsSuitable for: WB, Flow Cyt, IHC-Pmore details
Unsuitable for: ICC or IP -
Species reactivityReacts with: Human
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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.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
This product is a recombinant rabbit monoclonal antibody.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
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Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol, 0.05% BSA -
Concentration information loading... -
PurityProtein A purified
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ClonalityMonoclonal
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Clone numberE84
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IsotypeIgG
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Research areas
Associated products
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Alternative Products
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Compatible Secondaries
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Isotype control
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Positive Controls
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. |
Target
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FunctionInvolved 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 specificityUbiquitous, 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.
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Sequence similaritiesBelongs to the peptidase C14A family.
Contains 1 CARD domain. -
Developmental stageExpressed at low levels in fetal heart, at moderate levels in neonate heart, and at high levels in adult heart.
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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-ab32068KOWB.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-Untitled.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/pro-Caspase-9-Primary-antibodies-ab32068-1.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). -
![Western blot - Anti-Caspase-9 antibody [E84] (ab32068)](https://www.abcam.com/ps/products/32/ab32068/Images/ab32068_1.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
Datasheets and documents
References
This product has been referenced in:
- 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). Read more (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). Read more (PubMed: 30594603) »
