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Calreticulin Mouse Monoclonal Antibody [7B2]

Usd: 350

Specification

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Catalog# EM1701-61

Calreticulin Mouse Monoclonal Antibody [7B2]

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

Reactivity

  • Human

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Calreticulin Mouse Monoclonal Antibody [7B2]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Recombinant full length protein corresponding to Human Calretinin aa 1 to the C-terminus.

Species Reactivity

Human

Validated Applications

WB, IHC-P, IF-Cell, FC

Target Molecular Weight

Predicted band size: 48 kDa

Positive Control

HepG2 cell lysate, HeLa cell lysate, HL-60 cell lysate, HeLa , human thyroid gland tissue, human prostate cancer tissue, human placenta tissue, Jurkat.

Conjugation

unconjugated

Clone Number

7B2

RRID

AB_3068785

Product Features

Form

Liquid

Concentration

2 mg/mL.(The concentration of this product may be batch-dependent)

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

Isotype

IgG1

Purification Method

Protein G affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IF-Cell

  • 1:250

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

Target

Function

Calnexin and calregulin (also called calreticulin) are calcium-binding proteins that are localized to the endoplasmic reticulum, Calnexin to the membrane and calregulin to the lumen. Calnexin is a type I membrane protein that interacts with newly synthesized glycoproteins in the endoplasmic reticulum. It may play a role in assisting with protein assembly and in retaining unassembled protein subunits in the endoplasmic reticulum. Calregulin has both low- and high-affinity calcium-binding sites. Neither Calnexin nor calregulin contains the calcium-binding "E-F hand" motif found in calmodulins. Calnexin and calregulin are important for the maturation of glycoproteins in the endoplasmic reticulum and appear to bind many of the same proteins.

Background References

1. Nauseef W M et al. Calreticulin functions as a molecular chaperone in the biosynthesis of myeloperoxidase. J Biol Chem 270:4741-4747 (1995).

2. Holaska J M et al. Calreticulin is a receptor for nuclear export. J Cell Biol 152:127-140 (2001).

Sequence Similarity

Belongs to the calreticulin family.

Subcellular Location

Endoplasmic reticulum. Secreted. Cytosol.

UNIPROT

SwissProt: P27797 Human

Synonyms

Autoantigen RO antibody

CALR antibody

CALR protein antibody

CALR_HUMAN antibody

Calregulin antibody

Calreticulin antibody

cC1qR antibody

CRP55 antibody

CRT antibody

CRTC antibody

Expand

Images

  • Western blot analysis of Calreticulin on different lysates with Mouse anti-Calreticulin antibody (EM1701-61) at 1/5,000 dilution.<br /><br />Lane 1: HepG2 cell lysate (20 µg/Lane)<br />Lane 2: HeLa cell lysate (20 µg/Lane)<br />Lane 3: HL-60 cell lysate (20 µg/Lane)<br /><br />Predicted band size: 48 kDa<br />Observed band size: 55 kDa<br /><br />Exposure time: 1 minute;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1701-61) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Calreticulin on different lysates with Mouse anti-Calreticulin antibody (EM1701-61) at 1/5,000 dilution.

    Lane 1: HepG2 cell lysate (20 µg/Lane)
    Lane 2: HeLa cell lysate (20 µg/Lane)
    Lane 3: HL-60 cell lysate (20 µg/Lane)

    Predicted band size: 48 kDa
    Observed band size: 55 kDa

    Exposure time: 1 minute;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1701-61) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Calreticulin on different lysates with Mouse anti-Calreticulin antibody (EM1701-61) at 1/50,000 dilution.<br /><br />Lane 1: HeLa-si NT cell lysate<br />Lane 2: HeLa-si Calreticulin cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 48 kDa<br />Observed band size: 55 kDa<br /><br />Exposure time: 13 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1701-61) at 1/50,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of Calreticulin on different lysates with Mouse anti-Calreticulin antibody (EM1701-61) at 1/50,000 dilution.

    Lane 1: HeLa-si NT cell lysate
    Lane 2: HeLa-si Calreticulin cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 48 kDa
    Observed band size: 55 kDa

    Exposure time: 13 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1701-61) at 1/50,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells labeling Calreticulin with Mouse anti-Calreticulin antibody (EM1701-61) at 1/250 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-Calreticulin antibody (EM1701-61) at 1/250 dilution in 1% BSA in PBST 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.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling Calreticulin with Mouse anti-Calreticulin antibody (EM1701-61) at 1/250 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-Calreticulin antibody (EM1701-61) at 1/250 dilution in 1% BSA in PBST 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.

    beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human thyroid gland tissue using anti-Calreticulin antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human thyroid gland tissue using anti-Calreticulin antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue using anti-Calreticulin antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue using anti-Calreticulin antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-Calreticulin antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-Calreticulin antibody. Counter stained with hematoxylin.

Citation

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"