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Calpastatin Mouse Monoclonal Antibody [A6B1]

Usd: 350

Specification

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Catalog# HA600072

Calpastatin Mouse Monoclonal Antibody [A6B1]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

Calpastatin Mouse Monoclonal Antibody [A6B1]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Recombinant protein within human Calpastatin aa 501-708 / 708.

Species Reactivity

Human

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size: 120 kDa

Positive Control

A549 cell lysate, A431 cell lysate, Hela cell lysate, SiHa cell lysate A549, human esophagus tissue,.

Conjugation

unconjugated

Clone Number

A6B1

RRID

AB_3071689

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein G affinity purified.

Application Dilution

  • WB

  • 1:500

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:200

Target

Function

The protein encoded by this gene is an endogenous calpain (calcium-dependent cysteine protease) inhibitor. It consists of an N-terminal domain L and four repetitive calpain-inhibition domains (domains 1-4), and it is involved in the proteolysis of amyloid precursor protein. The calpain/calpastatin system is involved in numerous membrane fusion events, such as neural vesicle exocytosis and platelet and red-cell aggregation. The encoded protein is also thought to affect the expression levels of genes encoding structural or regulatory proteins. Alternatively spliced transcript variants encoding different isoforms have been described.

Background References

1. De Tullio R. et. al. Production and Purification of Recombinant Calpastatin. Methods Mol Biol. 2019

2. Averna M. et. al. Immunoblotting for Calpastatin Expression. Methods Mol Biol. 2019

Subcellular Location

Cytosol, endoplasmic reticulum, cytoplasm, membrane.

UNIPROT

SwissProt: P20810 Human

Synonyms

BS 17 antibody

Calpain inhibitor antibody

Calpastatin antibody

Cast antibody

Heart type calpastatin antibody

ICAL_HUMAN antibody

MGC9402 antibody

Sperm BS 17 component antibody

Sperm BS-17 component antibody

Images

  • Western blot analysis of on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA600072" style="font-weight: bold;text-decoration: underline;">HA600072</a>, 1/500) was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:20,000 dilution was used for 1 hour at room temperature.<br />Positive control:<br />Lane 1: A549 cell lysate<br />Lane 2: A431 cell lysate<br /><br />Predicted band size: 76.5 kDa<br />Observed band size: 120 kDa

    Western blot analysis of on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA600072, 1/500) was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 1 hour at room temperature.
    Positive control:
    Lane 1: A549 cell lysate
    Lane 2: A431 cell lysate

    Predicted band size: 76.5 kDa
    Observed band size: 120 kDa

  • Western blot analysis of Calpastatin on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/HA600072" style="font-weight: bold;text-decoration: underline;">HA600072</a>, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:20,000 dilution was used for 1 hour at room temperature.<br />Positive control:<br />Lane 1: Hela cell lysate<br />Lane 2: SiHa cell lysate<br /><br />Predicted band size: 76.5 kDa<br />Observed band size: 110-120 kDa

    Western blot analysis of Calpastatin on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (HA600072, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 1 hour at room temperature.
    Positive control:
    Lane 1: Hela cell lysate
    Lane 2: SiHa cell lysate

    Predicted band size: 76.5 kDa
    Observed band size: 110-120 kDa

  • Immunocytochemistry analysis of A549 cells labeling Calpastatin with Mouse anti-Calpastatin antibody (<a href="/products/HA600072" style="font-weight: bold;text-decoration: underline;">HA600072</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-Calpastatin antibody (<a href="/products/HA600072" style="font-weight: bold;text-decoration: underline;">HA600072</a>) at 1/100 dilution in 2% BSA overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of A549 cells labeling Calpastatin with Mouse anti-Calpastatin antibody (HA600072) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-Calpastatin antibody (HA600072) at 1/100 dilution in 2% BSA overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

  • Immunohistochemical analysis of paraffin-embedded human esophagus tissue using anti-Calpastatin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA600072" style="font-weight: bold;text-decoration: underline;">HA600072</a>, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human esophagus tissue using anti-Calpastatin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA600072, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of A549 cells labeling Calpastatin.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA600072" style="font-weight: bold;text-decoration: underline;">HA600072</a>, 1/1,000) (red) compared with Mouse IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Mouse IgG Secondary antibody (<a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of A549 cells labeling Calpastatin.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA600072, 1/1,000) (red) compared with Mouse IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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