Recombinant Anti-Caspase-6/CASP-6 antibody [EPR18043] (ab185645)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18043] to Caspase-6/CASP-6
- Suitable for: WB, IHC-P, IP, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Caspase-6/CASP-6 antibody [EPR18043]
See all Caspase-6/CASP-6 primary antibodies -
Description
Rabbit monoclonal [EPR18043] to Caspase-6/CASP-6 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, IP, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Jurkat, NIH/3T3, C6 and PC-12 whole cell lysates. IHC-P: Human colon, mouse brain and rat brain tissues. ICC/IF: Jurkat cells. IP: NIH/3T3 whole cell lysate treated with 1uM staurosporine 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.
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
EPR18043 -
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
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab185645 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 33, 11 kDa (predicted molecular weight: 33 kDa).
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IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
1/50.
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ICC/IF |
1/500.
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Notes |
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WB
1/1000. Detects a band of approximately 33, 11 kDa (predicted molecular weight: 33 kDa). |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
1/50. |
ICC/IF
1/500. |
Target
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Function
Involved in the activation cascade of caspases responsible for apoptosis execution. Cleaves poly(ADP-ribose) polymerase in vitro, as well as lamins. Overexpression promotes programmed cell death. -
Sequence similarities
Belongs to the peptidase C14A family. -
Post-translational
modificationsCleavages by caspase-3, caspase-8 or -10 generate the two active subunits. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 839 Human
- Entrez Gene: 12368 Mouse
- Entrez Gene: 83584 Rat
- Omim: 601532 Human
- SwissProt: P55212 Human
- SwissProt: O08738 Mouse
- SwissProt: O35397 Rat
- Unigene: 654616 Human
see all -
Alternative names
- Apoptotic protease Mch-2 antibody
- Apoptotic protease MCH2 antibody
- CASP-6 antibody
see all
Images
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All lanes : Anti-Caspase-6/CASP-6 antibody [EPR18043] (ab185645) at 1/1000 dilution
Lane 1 : Untreated Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate treated with 1uM staurosporine for 4 hours
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: 33 kDa
Observed band size: 11,33 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Caspase-6/CASP-6 antibody [EPR18043] (ab185645) at 1/1000 dilution
Lane 1 : Untreated NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate treated with 1uM staurosporine for 4 hours
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: 33 kDa
Observed band size: 11,33 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteThe predicted MW is 32kDa for mouse and rat full-length procaspase-6.
Blocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Caspase-6/CASP-6 antibody [EPR18043] (ab185645) at 1/1000 dilution
Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell 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: 33 kDa
Observed band size: 33 kDa
Exposure time: 1 minuteThe predicted MW is 32kDa for mouse and rat full-length procaspase-6.
Blocking/Dilution buffer: 5% NFDM/TBST.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling Caspase-6/CASP-6 with ab185645 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).
Confocal image showing both nuclear and cytoplasmic staining on Jurkat cells. The staining remained similar after treatment with staurosporine (1uM, 4 hours) as the antibody interacts with the subunit p11.
The nuclear counter stain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab185645 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution. -
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Caspase-6/CASP-6 with ab185645 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.
Cytoplasmic and nuclear staining on epithelial cells of human colon is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Mouse brain tissue labeling Caspase-6/CASP-6 with ab185645 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.
Cytoplasmic staining on neurons of mouse brain is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Rat brain tissue labeling Caspase-6/CASP-6 with ab185645 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.
Cytoplasmic staining on neurons of rat brain is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Caspase-6/CASP-6 was immunoprecipitated from 1mg of NIH/3T3 (Mouse embyro fibroblast cells) treated with 1uM staurosporine for 4 hours whole cell lysate with ab185645 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab185645 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1500 dilution.
Lane 1: NIH/3T3 treated with 1uM staurosporine for 4 hours whole cell lysate10 µg (Input). Lane 2: ab185645 IP in NIH/3T3 treated with 1uM staurosporine for 4 hours whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab185645 in NIH/3T3 treated with 1uM staurosporine for 4 hours whole cell lysate.
The cleaved Caspase-6/CASP-6 appears slightly larger than 11kDa. This fragment contains subunit p11 plus the internal propeptide.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (12)
ab185645 has been referenced in 12 publications.
- Chu H et al. Coronaviruses exploit a host cysteine-aspartic protease for replication. Nature 609:785-792 (2022). PubMed: 35922005
- Wu W et al. Caspase-Dependent Cleavage of DDX21 Suppresses Host Innate Immunity. mBio 12:e0100521 (2021). PubMed: 34125604
- Karvinen S et al. Estradiol deficiency and skeletal muscle apoptosis: Possible contribution of microRNAs. Exp Gerontol 147:111267 (2021). PubMed: 33548486
- Chen Y et al. Downregulation of phosphoglycerate mutase 5 improves microglial inflammasome activation after traumatic brain injury. Cell Death Discov 7:290 (2021). PubMed: 34642327
- He Y et al. CircZNF609 enhances hepatocellular carcinoma cell proliferation, metastasis, and stemness by activating the Hedgehog pathway through the regulation of miR-15a-5p/15b-5p and GLI2 expressions. Cell Death Dis 11:358 (2020). PubMed: 32398664