Tested Applications
| Positive WB detected in | HeLa cells, K-562 cells, MCF-7 cells |
| Positive IP detected in | K-562 cells |
| Positive IHC detected in | human lung cancer tissue Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0 |
| Positive IF/ICC detected in | HeLa cells |
| Positive FC (Intra) detected in | HeLa cells |
Recommended dilution
| Application | Dilution |
|---|---|
| Western Blot (WB) | WB : 1:2000-1:14000 |
| Immunoprecipitation (IP) | IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate |
| Immunohistochemistry (IHC) | IHC : 1:200-1:800 |
| Immunofluorescence (IF)/ICC | IF/ICC : 1:50-1:500 |
| Flow Cytometry (FC) (INTRA) | FC (INTRA) : 0.20 ug per 10^6 cells in a 100 µl suspension |
| It is recommended that this reagent should be titrated in each testing system to obtain optimal results. | |
| Sample-dependent, Check data in validation data gallery. | |
Product Information
12250-1-AP targets Calpastatin in WB, IHC, IF/ICC, FC (Intra), IP, ELISA applications and shows reactivity with human samples.
| Tested Reactivity | human |
| Cited Reactivity | human, mouse, rat |
| Host / Isotype | Rabbit / IgG |
| Class | Polyclonal |
| Type | Antibody |
| Immunogen |
CatNo: Ag2897 Product name: Recombinant human CAST protein Source: e coli.-derived, PGEX-4T Tag: GST Domain: 1-350 aa of BC013579 Sequence: MNPTETKAVKTEPEKKSQSTKPKSLPKQASDTGSNDAHNKKAVSRSAEQQPSEKSTEPKTKPQDMISAGGESVAGITAISGKPGDKKKEKKSLTPAVPVESKPDKPSGKSGMDAALDDLIDTLGGPEETEEENTTYTGPEVSDPMSSTYIEELGKREVTIPPKYRELLAKKEGITGPPADSSKPIGPDDAIDALSSDFTCGSPTAAGKKTEKEESTEVLKAQSAGTVRSAAPPQEKKRKVEKDTMSDQALEALSASLGTRQAEPELDLRSIKEVDEAKAKEEKLEKCGEDDETIPSEYRLKPATDKDGKPLLPEPEEKPKPRSESELIDELSEDFDRSECKEKPSKPTEK Predict reactive species |
| Full Name | calpastatin |
| Calculated Molecular Weight | 667 aa, 72 kDa |
| Observed Molecular Weight | 110 kDa |
| GenBank Accession Number | BC013579 |
| Gene Symbol | Calpastatin |
| Gene ID (NCBI) | 831 |
| RRID | AB_2071702 |
| Conjugate | Unconjugated |
| Form | Liquid |
| Purification Method | Antigen affinity purification |
| UNIPROT ID | P20810 |
| Storage Buffer | PBS with 0.02% sodium azide and 50% glycerol, pH 7.3. |
| Storage Conditions | Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage. 20ul sizes contain 0.1% BSA. |
Background Information
Calpastatin (CAST) is an endogenous protein that specifically inhibits the activity of calpains, a family of calcium-dependent proteases involved in numerous cellular processes. The CAST gene encodes calpastatin, which contains multiple calpain-inhibiting domains that bind to the active site of calpain, preventing it from performing its functions. Calpastatin is found in the cytosol and plays a key role in regulating intracellular proteolysis, with malfunctions in the calpain-calpastatin system linked to various diseases, including autoimmune disorders and neurodegenerative conditions.
Protocols
| Product Specific Protocols | |
|---|---|
| FC protocol for Calpastatin antibody 12250-1-AP | Download protocol |
| IF protocol for Calpastatin antibody 12250-1-AP | Download protocol |
| IHC protocol for Calpastatin antibody 12250-1-AP | Download protocol |
| IP protocol for Calpastatin antibody 12250-1-AP | Download protocol |
| WB protocol for Calpastatin antibody 12250-1-AP | Download protocol |
| Standard Protocols | |
|---|---|
| Click here to view our Standard Protocols |
Publications
What published studies show
The Calpain antibody (Cat# 12250-1-AP) has 6 total citations, all using WB (one also IF). Citations appear in Biomedicines, J Proteome Res, PeerJ, PLoS One, Ann Anat, and Oncotarget. Research fields span neurology (seizure/cognitive deficits, retinal neurons), virology (JAK-STAT1 signaling), cardiology (post-infarction sarcomere integrity), autophagy/acute pancreatitis, and oncology (chronic myeloid leukemia apoptosis). Species tested include rat, human, and mouse.
| Species | Application | Title |
|---|---|---|
Biomedicines Asiatic Acid Prevents Cognitive Deficits by Inhibiting Calpain Activation and Preserving Synaptic and Mitochondrial Function in Rats with Kainic Acid-Induced Seizure. | ||
J Proteome Res Quantitative Proteomic Analysis Reveals that Transmissible Gastroenteritis Virus (TGEV) Activates JAK-STAT1 Signaling Pathway. | ||
PeerJ Post-infarction KLHL40-mediated regulation of cardiac sarcomeric integrity and function. | ||
PLoS One Identification of autophagy-related key biomarkers in caerulein induced acute pancreatitis: In silico and in vivo study. | ||
Ann Anat Calpain2 but not calpain1 mediated by calpastatin following glutamate-induced regulated necrosis in rat retinal neurons. | ||
Oncotarget Decreased calpain activity in chronic myeloid leukemia impairs apoptosis by increasing survivin in myeloid progenitors and xiap1 in differentiating granulocytes. |
Reviews
What customers say
The single review for this antibody reports a 5-star rating from a Western Blot application on mouse heart lysates. The user noted a strong, predicted band, indicating reliable performance and specificity for the intended target. Overall, the feedback is positive, with the antibody delivering clear and expected results in WB.
The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.
Wei (Verified Customer) (03-12-2020) | strong and predicted band for western blot from heart lysates
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