Anti-Caspase-7 antibody [E22] (ab32522)
![Western blot - Anti-Caspase-7 antibody [E22] (ab32522)](https://www.abcam.com/ps/products/32/ab32522/Images/ab32522-409559-recombinant-anti-hmgb1-antibody-epr3506-western-blot-hela-lysates.jpg "Western blot - Anti-Caspase-7 antibody [E22] (ab32522)")
Key features and details
- Rabbit monoclonal [E22] to Caspase-7
- Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Related conjugates and formulations
Overview
-
Product name
Anti-Caspase-7 antibody [E22]
See all Caspase-7 primary antibodies -
Description
Rabbit monoclonal [E22] to Caspase-7 -
Host species
Rabbit -
Specificity
The antibody should recognize both pro-form and p20 cleaved-form. The antibody does not cross-react with other Caspase family members. -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: Jurkat and HeLa whole cell lysate (ab150035). IHC-P: Human skin cancer tissue. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells. IP: Jurkat cell lysates
-
General notes
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
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
E22 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Isotype control
-
KO cell lines
-
KO cell lysates
-
Positive Controls
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32522 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) |
1/1000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
|
| IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
|
| ICC/IF |
1/100.
|
|
| IP | (1) |
1/20.
|
| Flow Cyt (Intra) |
1/250.
|
| Notes |
|---|
|
WB
1/1000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa). |
|
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
|
ICC/IF
1/100. |
|
IP
1/20. |
|
Flow Cyt (Intra)
1/250. |
Target
-
Function
Involved in the activation cascade of caspases responsible for apoptosis execution. Cleaves and activates sterol regulatory element binding proteins (SREBPs). Proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp-
-Gly-217' bond. Overexpression promotes programmed cell death. -
Tissue specificity
Highly expressed in lung, skeletal muscle, liver, kidney, spleen and heart, and moderately in testis. No expression in the brain. -
Sequence similarities
Belongs to the peptidase C14A family. -
Post-translational
modificationsCleavages by granzyme B or caspase-10 generate the two active subunits. Propeptide domains can also be cleaved efficiently by caspase-3. Active heterodimers between the small subunit of caspase-7 and the large subunit of caspase-3, and vice versa, also occur. -
Cellular localization
Cytoplasm. - Information by UniProt
-
Database links
- Entrez Gene: 840 Human
- Omim: 601761 Human
- SwissProt: P55210 Human
- Unigene: 9216 Human
-
Alternative names
- Apoptotic protease Mch-3 antibody
- Apoptotic protease MCH3 antibody
- CASP-7 antibody
see all
Images
-
Western blot - Anti-Caspase-7 antibody [E22] (ab32522)All lanes : Anti-Caspase-7 antibody [E22] (ab32522) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CASP7 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab32522 observed at 38 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab32522 was shown to react with pro Caspase-7 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265777 (knockout cell lysate ab257380) was used. Wild-type HeLa and CASP7 knockout HeLa cell lysates were subjected to SDS-PAGE. ab32522 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
-
![Western blot - Anti-Caspase-7 antibody [E22] (ab32522)](https://www.abcam.com/ps/products/32/ab32522/Images/ab32522-45103-anti-caspase-7-antibody-e22-western-blot.gif)
Western blot - Anti-Caspase-7 antibody [E22] (ab32522)Anti-Caspase-7 antibody [E22] (ab32522) at 1/1000 dilution + Jurkat cell lysate
Predicted band size: 34 kDa
Observed band size: 34 kDa -
![Western blot - Anti-Caspase-7 antibody [E22] (ab32522)](https://www.abcam.com/ps/products/32/ab32522/Images/ab32522-283568-anti-caspase-7-antibody-e22-western-blot.jpg)
Western blot - Anti-Caspase-7 antibody [E22] (ab32522)Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: Wild type HAP1 + Staurosporine ab120056 whole cell lysate (20 µg)
Lane 3: CASP7 knockout HAP1 whole cell lysate (20 µg)
Lane 4: CASP7 + Staurosporine knockout HAP1 whole cell lysate (20 µg)
Lane 5: HeLa whole cell lysate (20 µg)
Lane 6: HeLa + Staurosporine whole cell lysate (20 µg)Lanes 1 - 6: Merged signal (red and green). Green - ab32522 observed at 38 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab32522 was shown to specifically react with HAP1 + Staurosporine when HAP1 + Staurosporine knockout samples were used. Wild-type and HAP1 + Staurosporine knockout samples were subjected to SDS-PAGE. Ab32522 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 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.
-
![Immunocytochemistry/ Immunofluorescence - Anti-Caspase-7 antibody [E22] (ab32522)](https://www.abcam.com/ps/products/32/ab32522/Images/ab32522-45101-anti-caspase-7-antibody-e22-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Caspase-7 antibody [E22] (ab32522)Immunofluorescent staining of HeLa cells using ab32522 at 1:100 dilution.
-
![Flow Cytometry (Intracellular) - Anti-Caspase-7 antibody [E22] (ab32522)](https://www.abcam.com/ps/products/32/ab32522/Images/ab32522-280379-anti-caspase-7-antibody-e22-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-Caspase-7 antibody [E22] (ab32522)Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Caspase-7 (red) with ab32522 at a 1/250 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluorr® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-7 antibody [E22] (ab32522)](https://www.abcam.com/ps/products/32/ab32522/Images/ab32522-45102-anti-caspase-7-antibody-e22-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-7 antibody [E22] (ab32522)Immunohistochemical analysis of paraffin embedded human skin cancer tissue using ab32522 at 1:50 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
-
![Immunoprecipitation - Anti-Caspase-7 antibody [E22] (ab32522)](https://www.abcam.com/ps/products/32/ab32522/Images/ab32522-8-anti-caspase-7-antibody-e22-immunoprecipitation-jurkat-human.jpg)
Immunoprecipitation - Anti-Caspase-7 antibody [E22] (ab32522)Purified ab32522 at 1/20 dilution (1µg) immunoprecipitating Caspase-7 in Jurkat whole cell lysate.
Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10µg
Lane 2 (+): ab32522 + Jurkat whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32522 in Jurkat whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 34 kDa
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
-
Datasheet download
References (22)
ab32522 has been referenced in 22 publications.
- Zhao X et al. Casein Kinase 2-Interacting Protein-1 Alleviates High Glucose-Reduced Autophagy, Oxidative Stress, and Apoptosis in Retinal Pigment Epithelial Cells via Activating the p62/KEAP1/NRF2 Signaling Pathway. J Ophthalmol 2021:6694050 (2021). PubMed: 33628480
- Gao Z et al. FK506-binding protein 5 promotes the progression of papillary thyroid carcinoma. J Int Med Res 49:3000605211008325 (2021). PubMed: 33906532
- Chang Y et al. MIR205HG facilitates carcinogenesis of lung squamous cell carcinoma in vitro revealed by long noncoding RNA profiling. Acta Biochim Biophys Sin (Shanghai) 52:371-381 (2020). PubMed: 32188965
- Albahde MAH et al. CBL0137 administration suppresses human hepatocellular carcinoma cells proliferation and induces apoptosis associated with multiple cell death related proteins. Neoplasma 67:547-556 (2020). PubMed: 32202904
- Wu SQ et al. A novel role for Livin in the response to ultraviolet B radiation and pterygium development. Int J Mol Med 45:1103-1111 (2020). PubMed: 32124942
