Recombinant Anti-Caspase-3 antibody [E87] - BSA and Azide free (ab197202)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E87] to Caspase-3 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), IP, IHC-P, WB, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Caspase-3 antibody [E87] - BSA and Azide free
See all Caspase-3 primary antibodies -
Description
Rabbit monoclonal [E87] to Caspase-3 - BSA and Azide free -
Host species
Rabbit -
Specificity
This antibody is specific for the pro form and the p17 cleaved form of human Caspase-3.
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Tested applications
Suitable for: Flow Cyt (Intra), IP, IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Human
Does not react with: Mouse -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Wild-type HAP1 whole cell lysate. IHC-P: Human tonsil and cervical carcinoma tissue. ICC/IF: Jurkat cells and wild-type HAP1 cells. Flow Cyt (intra): HeLa and Ramos cells. IP: HeLa whole cell lysate.
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General notes
ab197202 is the carrier-free version of ab32351.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
E87 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Biotin Anti-Caspase-3 antibody [E87] (ab195905)
- PE Anti-Caspase-3 antibody [E87] (ab305665)
- APC Anti-Caspase-3 antibody [E87] (ab305666)
- HRP Anti-Caspase-3 antibody [E87] (ab305667)
- Alexa Fluor® 488 Anti-Caspase-3 antibody [E87] (ab309678)
- Alexa Fluor® 647 Anti-Caspase-3 antibody [E87] (ab310040)
- Alexa Fluor® 594 Anti-Caspase-3 antibody [E87] (ab310425)
- Alexa Fluor® 555 Anti-Caspase-3 antibody [E87] (ab311951)
- Alexa Fluor® 568 Anti-Caspase-3 antibody [E87] (ab312424)
- Anti-Caspase-3 antibody [E87] (ab32351)
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Compatible Secondaries
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Conjugation kits
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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 ab197202 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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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 35 kDa (predicted molecular weight: 32 kDa).
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ICC/IF |
Use at an assay dependent concentration.
For unpurified, use 1/25 dilution. |
Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. Detects a band of approximately 35 kDa (predicted molecular weight: 32 kDa). |
ICC/IF
Use at an assay dependent concentration. For unpurified, use 1/25 dilution. |
Target
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Function
Involved in the activation cascade of caspases responsible for apoptosis execution. At the onset of apoptosis it proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp-
-Gly-217' bond. Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain. Cleaves and activates caspase-6, -7 and -9. Involved in the cleavage of huntingtin. -
Tissue specificity
Highly expressed in lung, spleen, heart, liver and kidney. Moderate levels in brain and skeletal muscle, and low in testis. Also found in many cell lines, highest expression in cells of the immune system. -
Sequence similarities
Belongs to the peptidase C14A family. -
Post-translational
modificationsCleavage by granzyme B, caspase-6, caspase-8 and caspase-10 generates the two active subunits. Additional processing of the propeptides is likely due to the autocatalytic activity of the activated protease. Active heterodimers between the small subunit of caspase-7 protease and the large subunit of caspase-3 also occur and vice versa.
S-nitrosylated on its catalytic site cysteine in unstimulated human cell lines and denitrosylated upon activation of the Fas apoptotic pathway, associated with an increase in intracellular caspase activity. Fas therefore activates caspase-3 not only by inducing the cleavage of the caspase zymogen to its active subunits, but also by stimulating the denitrosylation of its active site thiol. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 836 Human
- Omim: 600636 Human
- SwissProt: P42574 Human
- Unigene: 141125 Human
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Alternative names
- A830040C14Rik antibody
- Apopain antibody
- CASP 3 antibody
see all
Images
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All lanes : Anti-Caspase-3 antibody [E87] (ab32351) at 1/5000 dilution
Lane 1 : DMSO control wild-type HAP1 whole cell lysate
Lane 2 : Staurosporine treated wild-type HAP1 whole cell lysate
Lane 3 : DMSO control CASP3 knockout HAP1 whole cell lysate
Lane 4 : Staurosporine treated CASP3 knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 32 kDaLanes 1 - 4: Merged signal (red and green). Green - ab32351 observed at 31 kDa. Red - loading control, ab130007, observed at 130 kDa.
ab32351 was shown to recognize Caspase 3 in wild-type HAP1 cells as signal was lost at the expected MW in HAP1 Staurosporine Treated (CASP3) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and HAP1 Staurosporine Treated (CASP3) knockout samples were subjected to SDS-PAGE. ab32351 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 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 1/20000 dilution for 1 hour at room temperature before imaging.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32351).
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This data was developed using the same antibody clone in a different buffer formulation (ab32351). ab32351 staining Caspase-3 in wild-type Hap1 cells (top panel) and CASP3 knockout Hap1 cells (bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab32351 at 1μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8). -
This IHC data was generated using the same anti-Caspase 3 antibody clone, E87, in a different buffer formulation (cat# ab32351).
Immunohistochemical staining of paraffin embedded human tonsil with purified ab32351 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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ab32351 (purified) at 1/50 immunoprecipitating Cullin 1 in 10 µg HeLa whole cell lysate (Lanes 1 and 2, observed at 35 kDa). Lane 3 - PBS. For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32351).
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Immunofluorescence staining of Jurkat cells with purified ab32351 at a working dilution of 1/500, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab32351 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32351).
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Overlay histogram showing Ramos cells fixed in 4% PFA and stained with purified ab32351 at a dilution of 1 in 180 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32351).
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Overlay histogram showing HeLa cells stained with unpurfied ab32351 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32351, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluorr® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730, 0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32351).
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (51)
ab197202 has been referenced in 51 publications.
- Almasoud HA et al. Dose-Dependent Variation in Anticancer Activity of Hexane and Chloroform Extracts of Field Horsetail Plant on Human Hepatocarcinoma Cells. Biomed Res Int 2022:5778411 (2022). PubMed: 35789647
- Liao Y et al. miR‑302d‑3p regulates the viability, migration and apoptosis of breast cancer cells through regulating the TMBIM6‑mediated ERK signaling pathway. Mol Med Rep 24:N/A (2021). PubMed: 34651659
- Zhang Y et al. Inhibition of miR-15a-5p Promotes the Chemoresistance to Pirarubicin in Hepatocellular Carcinoma via Targeting eIF4E. Comput Math Methods Med 2021:6468405 (2021). PubMed: 34812269
- Wang M et al. Dynamic study into autophagy and apoptosis during orthodontic tooth movement. Exp Ther Med 21:430 (2021). PubMed: 33747169
- Mao L et al. Combination of oncolytic adenovirus targeting SATB1 and docetaxel for the treatment of castration-resistant prostate cancer. J Cancer 12:1846-1852 (2021). PubMed: 33613773