Recombinant Anti-Caspase-9 antibody [EPR18108] (ab185719)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18108] to Caspase-9
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Mouse, Human
Overview
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Product name
Anti-Caspase-9 antibody [EPR18108]
See all Caspase-9 primary antibodies -
Description
Rabbit monoclonal [EPR18108] to Caspase-9 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IPmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Recombinant fragment within Human Caspase-9 aa 100-350. The exact sequence is proprietary.
Database link: P55211 -
Positive control
- WB: HeLa whole cell lysates untreated and treated with staurosporine 1uM for 4 hours; NIH/3T3 whole cell lysates untreated and treated with staurosporine1uM for 4 hours; Human fetal brain, fetal heart, fetal kidney and fetal liver and pancreas lysates; Mouse heart and kidney lysates. IP: HeLa whole cell lysate treated with staurosporine 1uM for 4 hours.
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General notes
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18108 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Assay kits
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab185719 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 |
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WB | 1/1000. Detects a band of approximately 46, 37, 35 kDa (predicted molecular weight: 46 kDa). | |
IP | 1/40. |
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
- Entrez Gene: 12371 Mouse
- Omim: 602234 Human
- SwissProt: P55211 Human
- SwissProt: Q8C3Q9 Mouse
- Unigene: 329502 Human
- Unigene: 88829 Mouse
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Alternative names
- APAF-3 antibody
- APAF3 antibody
- Apoptosis related cysteine peptidase antibody
see all
Images
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Caspase-9 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Jurkat cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab185719 observed at 46 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab185719 was shown to specifically react with Caspase-9 when Caspase-9 knockout samples were used. Wild-type and Caspase-9 knockout samples were subjected to SDS-PAGE. ab185719 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging. -
All lanes : Anti-Caspase-9 antibody [EPR18108] (ab185719) at 1/10000 dilution
Lane 1 : Untreated HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa whole cell lysate treated with staurosporine 1uM for 4 hours
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 46 kDa
Observed band size: 35,37,46 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Caspase-9 antibody [EPR18108] (ab185719) at 1/2500 dilution
Lane 1 : Untreated NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate
Lane 2 : NIH/3T3 whole cell lysate treated with staurosporine 1uM for 4 hours
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 46 kDa
Observed band size: 37,39,49 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Caspase-9 antibody [EPR18108] (ab185719) at 1/1000 dilution
Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 46 kDa
Observed band size: 46 kDa
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Caspase-9 antibody [EPR18108] (ab185719) at 1/5000 dilution
Lane 1 : Human pancreas lysate
Lane 2 : Human fetal liver lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 46 kDa
Observed band size: 46 kDa
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Caspase-9 antibody [EPR18108] (ab185719) at 1/1000 dilution
Lane 1 : Mouse heart lysate
Lane 2 : Mouse kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 46 kDa
Observed band size: 37,39,49 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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Caspase-9 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate treated with staurosporine 1uM for 4 hours with ab185719 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab185719 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: HeLa whole cell lysate treated with staurosporine 10 µg (Input).
Lane 2: ab185719 IP in HeLa whole cell lysate treated with staurosporine.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab185719 in HeLa whole cell lysate treated with staurosporine.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Protocols
Datasheets and documents
Certificate of Compliance
References (5)
ab185719 has been referenced in 5 publications.
- Huang Y et al. Dehydrocorydaline inhibits the tumorigenesis of breast cancer MDA-MB-231 cells. Mol Med Rep 22:43-50 (2020). PubMed: 32377708
- Yagensky O et al. Increased expression of heme-binding protein 1 early in Alzheimer's disease is linked to neurotoxicity. Elife 8:N/A (2019). PubMed: 31453805
- Wang X et al. Silence of lncRNA ANRIL represses cell growth and promotes apoptosis in retinoblastoma cells through regulating miR-99a and c-Myc. Artif Cells Nanomed Biotechnol 47:2265-2273 (2019). PubMed: 31184221
- Zhong X et al. Downregulation of microRNA-34a inhibits oxidized low-density lipoprotein-induced apoptosis and oxidative stress in human umbilical vein endothelial cells. Int J Mol Med 42:1134-1144 (2018). PubMed: 29750293
- Luff SA et al. Role of p53 and transcription-independent p53-induced apoptosis in shear-stimulated megakaryocytic maturation, particle generation, and platelet biogenesis. PLoS One 13:e0203991 (2018). PubMed: 30231080