Calpastatin Recombinant monoclonal antibody

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• Calpastatin Recombinant monoclonal antibody (86664-3-RR)

• KD/KO Validated

Calpastatin Uni-rAb^® Recombinant Antibody for WB, IHC, IF/ICC, ELISA

Cat No. 86664-3-RR

Clone No.251652G2

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, IHC, IF/ICC, ELISA

Synonyms

CAST, Calpain inhibitor, Sperm BS-17 component

• 

  WB analysis of HeLa using 86664-3-RR

  WB result of Calpastatin antibody (86664-3-RR; 1:10000; incubated at room temperature for 1.5 hours) with sh-Control and sh-Calpastatin transfected HeLa cells.

  This validation data has been generated in-house and reviewed by Proteintech scientists. Find out more on our antibody validation policy
• 

  WB analysis using 86664-3-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 86664-3-RR (CAST antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

  This validation data has been generated in-house and reviewed by Proteintech scientists. Find out more on our antibody validation policy
• 

  IHC staining of human heart using 86664-3-RR

  Immunohistochemical analysis of paraffin-embedded human heart tissue slide using 86664-3-RR (Calpastatin antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  This validation data has been generated in-house and reviewed by Proteintech scientists. Find out more on our antibody validation policy
• 

  IHC staining of human heart using 86664-3-RR

  Immunohistochemical analysis of paraffin-embedded human heart tissue slide using 86664-3-RR (Calpastatin antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  This validation data has been generated in-house and reviewed by Proteintech scientists. Find out more on our antibody validation policy
• 

  IF Staining of HeLa using 86664-3-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed HeLa cells using Calpastatin antibody (86664-3-RR, Clone: 251652G2 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2).

  This validation data has been generated in-house and reviewed by Proteintech scientists. Find out more on our antibody validation policy
• 

  Affinity and Kinetic Characterization of 86664-3-RR

  Biolayer interferometry (BLl) kinetic assays of 86664-3-RR against Human Calpastatin were performed. The affinity constant is 6.39 nM.

  This validation data has been generated in-house and reviewed by Proteintech scientists. Find out more on our antibody validation policy

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Tested Applications

• Positive WB detected in: HeLa cells, K-562 cells, MDA-MB-231 cells, A549 cells, HaCaT cells
• Positive IHC detected in: human heart 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

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800

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

86664-3-RR targets Calpastatin in WB, IHC, IF/ICC, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• 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

• Full Name: calpastatin
• Calculated Molecular Weight: 667 aa, 72 kDa
• Observed Molecular Weight: 110-130 kDa
• GenBank Accession Number: BC013579
• Gene Symbol: Calpastatin
• Gene ID (NCBI): 831
• RRID: AB_3745040
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A 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 -20^oC 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.