Validation Data Gallery

Filter: AllWBIPIHCIF/ICCFC (Intra)

WB analysis using 12250-1-AP
Various lysates were subjected to SDS PAGE followed by western blot with 12250-1-AP (Calpastatin antibody) at dilution of 1:7000 incubated at room temperature for 1.5 hours.
WB analysis of MCF-7 using 12250-1-AP
MCF7 cells were subjected to SDS PAGE followed by western blot with 12250-1-AP (Calpastatin antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.
WB analysis of K-562 using 12250-1-AP
K-562 cells were subjected to SDS PAGE followed by western blot with 12250-1-AP (Calpastatin antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.
IP experiment of K-562 using 12250-1-AP
IP result of anti-Calpastatin (IP:12250-1-AP, 4ug; Detection:12250-1-AP 1:1000) with K-562 cells lysate 4000ug.
IHC staining of human lung cancer using 12250-1-AP
Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 12250-1-AP (Calpastatin antibody) at dilution of 1:400 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of human lung cancer using 12250-1-AP
Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 12250-1-AP (Calpastatin antibody) at dilution of 1:400 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IF Staining of HeLa using 12250-1-AP
Immunofluorescent analysis of (-20°C Methanol) fixed HeLa cells using Calpastatin antibody (12250-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).
FC experiment of HeLa using 12250-1-AP
1X10^6 HeLa cells were intracellularly stained with 0.2 ug Anti-Human Calpastatin (12250-1-AP) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.2 ug Control Antibody. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

Calpastatin Polyclonal antibody

Calpastatin Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Cat No. 12250-1-AP

Publications(4)

Host / Isotype

Rabbit / IgG

Reactivity

human and More (1)

Applications

WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Synonyms

CAST, Calpain inhibitor, Sperm BS-17 component

Formulation: PBS and Azide

PBS and Azide

Conjugate: Unconjugated

Unconjugated

Size/Concentration:

-/ -

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Overview

Tested Applications
Product Information
Publications (4)
Protocols

データシート［PDF］を印刷

MSDS

Tested Applications

Positive WB detected in	HeLa cells, MCF-7 cells, K-562 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.

Published Applications

WB	See 4 publications below
IF	See 1 publications below

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, 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 相同性解析による交差性が予測される生物種
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
Form	Liquid
Purification Method	Antigen affinity purification
UNIPROT ID	P20810
Storage Buffer	PBS with 0.02% sodium azide and 50% glycerol{{ptg:BufferTemp}}7.3
Storage Conditions	Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

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
WB protocol for Calpastatin antibody 12250-1-AP	Download protocol
IHC protocol for Calpastatin antibody 12250-1-AP	Download protocol
IF protocol for Calpastatin antibody 12250-1-AP	Download protocol
IP protocol for Calpastatin antibody 12250-1-AP	Download protocol
FC protocol for Calpastatin antibody 12250-1-AP	Download protocol

Standard Protocols
Click here to view our Standard Protocols

Publications

Species	Application	Title
human	WB	J Proteome Res Quantitative Proteomic Analysis Reveals that Transmissible Gastroenteritis Virus (TGEV) Activates JAK-STAT1 Signaling Pathway. Authors - Kang An View Article
rat	WB	Biomedicines Asiatic Acid Prevents Cognitive Deficits by Inhibiting Calpain Activation and Preserving Synaptic and Mitochondrial Function in Rats with Kainic Acid-Induced Seizure. Authors - Cheng-Wei Lu View Article
rat	WB,IF	Ann Anat Calpain2 but not calpain1 mediated by calpastatin following glutamate-induced regulated necrosis in rat retinal neurons. Authors - Shuchao Wang View Article
human	WB	Oncotarget Decreased calpain activity in chronic myeloid leukemia impairs apoptosis by increasing survivin in myeloid progenitors and xiap1 in differentiating granulocytes. Authors - Weiqi Huang View Article

Target

Host

Species Reactivity

Species Reactivity

Conjugate

Conjugate

Validation Data Gallery

WB analysis using 12250-1-AP

WB analysis of MCF-7 using 12250-1-AP

WB analysis of K-562 using 12250-1-AP

IP experiment of K-562 using 12250-1-AP

IHC staining of human lung cancer using 12250-1-AP

IHC staining of human lung cancer using 12250-1-AP

IF Staining of HeLa using 12250-1-AP

FC experiment of HeLa using 12250-1-AP