Calpastatin Recombinant monoclonal antibody

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

Cat No. 86664-3-RR

Clone No.251652G2

Review Product

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, IHC, IF/ICC, ELISA

Synonyms

CAST, Calpain inhibitor, Sperm BS-17 component

Formulation: PBS, Azide, Glycerol

PBS, Azide, Glycerol

PBS Only

Conjugate: Unconjugated

Unconjugated

Size/Concentration:

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-/ -

Freight/Packing: -

Quantity

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Filter: AllWBIHCIF/ICCAffinity

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

Overview

Tested Applications
Product Information
Protocols

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MSDS SDS

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

Application	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 Predict reactive species
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.

Protocols

Product Specific Protocols
IF protocol for Calpastatin antibody 86664-3-RR	Download protocol
IHC protocol for Calpastatin antibody 86664-3-RR	Download protocol
WB protocol for Calpastatin antibody 86664-3-RR	Download protocol