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Calreticulin Recombinant Rabbit Monoclonal Antibody [SU37-03] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750156

Calreticulin Recombinant Rabbit Monoclonal Antibody [SU37-03] - BSA and Azide free

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

Calreticulin Recombinant Rabbit Monoclonal Antibody [SU37-03] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Calreticulin aa 40-89 / 417.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Cell, FC, IP

Molecular Weight

Predicted band size: 48 kDa

Positive Control

HepG2 cell lysate, HeLa cell lysate, HL-60 cell lysate, C2C12 cell lysate, C6 cell lysate, Mouse liver tissue lysate, Rat liver tissue lysate, human liver tissue, human liver carcinoma tissue, HeLa.

Conjugation

unconjugated

Clone Number

SU37-03

Reactivity Data

WB IHC-P IF-Cell FC IP
Human
Tested Tested
Tested Tested
Tested Tested
Tested Tested
Tested Tested
Mouse
Tested Tested
Rat
Tested Tested
Not Recommended
Not Recommended

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:50,000

  • IHC-P

  • 1:5,000

  • IF-Cell

  • 1:5,000

  • FC

  • 1:2,000

  • IP

  • 1-2μg/sample

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. Kojima Y et al. Cyclin-dependent kinase inhibitor 2B regulates efferocytosis and atherosclerosis. J Clin Invest 124:1083-97 (2014).

2. Angelova AL et al. Complementary induction of immunogenic cell death by oncolytic parvovirus H-1PV and gemcitabine in pancreatic cancer. J Virol 88:5263-76 (2014).

Sequence Similarity

Belongs to the calreticulin family.

Subcellular Location

Endoplasmic reticulum lumen, Cytoplasm, Secreted, Cell surface, Sarcoplasmic reticulum lumen, nucleus.

UNIPROT

SwissProt: P27797 Human

SwissProt: P14211 Mouse

SwissProt: P18418 Rat

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

  • Application: Immunocytochemistry (IF-cell)<br /><br />Species: Human<br />Sample: HeLa (Human cervix adenocarcinoma epithelial cell)<br /><br />Fixation: Ice cold 100% methanol, 5 minutes at room temperature.<br />Permeabilization: /<br /><br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.<br />Antibody dilution buffer: 1% BSA in PBST.<br />Primary antibody: <a href="/products/HA750156" style="font-weight: bold;text-decoration: underline;">HA750156</a>, 1/5,000, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 45 minutes at room temperature.<br /><br />Counterstain: Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, Red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).

    Application: Immunocytochemistry (IF-cell)

    Species: Human
    Sample: HeLa (Human cervix adenocarcinoma epithelial cell)

    Fixation: Ice cold 100% methanol, 5 minutes at room temperature.
    Permeabilization: /

    Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
    Antibody dilution buffer: 1% BSA in PBST.
    Primary antibody: HA750156, 1/5,000, overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.

    Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).

  • Western blot analysis of Calreticulin on different lysates with Rabbit anti-Calreticulin antibody (<a href="/products/HA750156" style="font-weight: bold;text-decoration: underline;">HA750156</a>) at 1/50,000 dilution and competitor's antibody at 1/5,000 dilution.<br /><br />Lane 1: HepG2 cell lysate<br />Lane 2: HeLa cell lysate<br />Lane 3: HL-60 cell lysate<br />Lane 4: C2C12 cell lysate<br />Lane 5: C6 cell lysate<br />Lane 6: Mouse liver tissue lysate<br />Lane 7: Rat liver tissue 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: 1 minute 2 seconds; ECL: K1801;<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 (<a href="/products/HA750156" style="font-weight: bold;text-decoration: underline;">HA750156</a>) at 1/50,000 dilution and competitor's antibody at 1/5,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Calreticulin on different lysates with Rabbit anti-Calreticulin antibody (HA750156) at 1/50,000 dilution and competitor's antibody at 1/5,000 dilution.

    Lane 1: HepG2 cell lysate
    Lane 2: HeLa cell lysate
    Lane 3: HL-60 cell lysate
    Lane 4: C2C12 cell lysate
    Lane 5: C6 cell lysate
    Lane 6: Mouse liver tissue lysate
    Lane 7: Rat liver tissue lysate

    Lysates/proteins at 10 µg/Lane.

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

    Exposure time: 1 minute 2 seconds; ECL: K1801;

    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 (HA750156) at 1/50,000 dilution and competitor's antibody at 1/5,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) 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 Rabbit anti-Calreticulin antibody (<a href="/products/HA750156" style="font-weight: bold;text-decoration: underline;">HA750156</a>) at 1/50,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate (10 µg/Lane)<br />Lane 2: HAP1-Calreticulin KD cell lysate (10 µg/Lane)<br /><br />Predicted band size: 48 kDa<br />Observed band size: 55 kDa<br /><br />Exposure time: 30 seconds; ECL: K1801;<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 (<a href="/products/HA750156" style="font-weight: bold;text-decoration: underline;">HA750156</a>) at 1/50,000 dilution was used in <a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a> at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</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 Rabbit anti-Calreticulin antibody (HA750156) at 1/50,000 dilution.

    Lane 1: HAP1-parental cell lysate (10 µg/Lane)
    Lane 2: HAP1-Calreticulin KD cell lysate (10 µg/Lane)

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

    Exposure time: 30 seconds; ECL: K1801;
    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 (HA750156) at 1/50,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) 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 Rabbit anti-Calreticulin antibody (<a href="/products/HA750156" style="font-weight: bold;text-decoration: underline;">HA750156</a>) at 1/50,000 dilution.<br /><br />Lane 1: HeLa-si NT cell lysate (10 µg/Lane)<br />Lane 2: HeLa-si Calreticulin cell lysate (10 µg/Lane)<br /><br />Predicted band size: 48 kDa<br />Observed band size: 55 kDa<br /><br />Exposure time: 13 seconds; ECL: K1801;<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 (<a href="/products/HA750156" style="font-weight: bold;text-decoration: underline;">HA750156</a>) at 1/50,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</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 Rabbit anti-Calreticulin antibody (HA750156) at 1/50,000 dilution.

    Lane 1: HeLa-si NT cell lysate (10 µg/Lane)
    Lane 2: HeLa-si Calreticulin cell lysate (10 µg/Lane)

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

    Exposure time: 13 seconds; ECL: K1801;
    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 (HA750156) at 1/50,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Calreticulin antibody (<a href="/products/HA750156" style="font-weight: bold;text-decoration: underline;">HA750156</a>) at 1/5,000 dilution.<br /><br />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 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750156" style="font-weight: bold;text-decoration: underline;">HA750156</a>) at 1/5,000 dilution for 1 hour 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 liver tissue with Rabbit anti-Calreticulin antibody (HA750156) at 1/5,000 dilution.

    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 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750156) at 1/5,000 dilution for 1 hour 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 liver carcinoma tissue with Rabbit anti-Calreticulin antibody (<a href="/products/HA750156" style="font-weight: bold;text-decoration: underline;">HA750156</a>) at 1/5,000 dilution.<br /><br />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 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750156" style="font-weight: bold;text-decoration: underline;">HA750156</a>) at 1/5,000 dilution for 1 hour 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 liver carcinoma tissue with Rabbit anti-Calreticulin antibody (HA750156) at 1/5,000 dilution.

    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 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750156) at 1/5,000 dilution for 1 hour 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.

  • Application: Immunocytochemistry (IF-cell)<br /><br />Species: Human<br />Sample: HeLa (Human cervix adenocarcinoma epithelial cell) treated with 200nM PMA for 30 minutes<br /><br />Fixation: Ice cold 100% methanol, 5 minutes at room temperature.<br />Permeabilization: /<br /><br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.<br />Antibody dilution buffer: 1% BSA in PBST.<br />Primary antibody: <a href="/products/HA750156" style="font-weight: bold;text-decoration: underline;">HA750156</a>, 1/5,000, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 45 minutes at room temperature.<br /><br />Counterstain: Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, Red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).

    Application: Immunocytochemistry (IF-cell)

    Species: Human
    Sample: HeLa (Human cervix adenocarcinoma epithelial cell) treated with 200nM PMA for 30 minutes

    Fixation: Ice cold 100% methanol, 5 minutes at room temperature.
    Permeabilization: /

    Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
    Antibody dilution buffer: 1% BSA in PBST.
    Primary antibody: HA750156, 1/5,000, overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.

    Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).

  • Flow cytometric analysis of HeLa cells labeling Calreticulin.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA750156" style="font-weight: bold;text-decoration: underline;">HA750156</a>, 1/2,000) (red) compared with Rabbit 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-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</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 HeLa cells labeling Calreticulin.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA750156, 1/2,000) (red) compared with Rabbit 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-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Calreticulin was immunoprecipitated in 0.2mg HeLa cell lysate with <a href="/products/HA750156" style="font-weight: bold;text-decoration: underline;">HA750156</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA750156" style="font-weight: bold;text-decoration: underline;">HA750156</a> at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HeLa cell lysate (input)<br />Lane 2: Rabbit IgG instead of <a href="/products/HA750156" style="font-weight: bold;text-decoration: underline;">HA750156</a> in HeLa cell lysate<br />Lane 3: <a href="/products/HA750156" style="font-weight: bold;text-decoration: underline;">HA750156</a> IP in HeLa cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 24 seconds

    Calreticulin was immunoprecipitated in 0.2mg HeLa cell lysate with HA750156 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA750156 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa cell lysate (input)
    Lane 2: Rabbit IgG instead of HA750156 in HeLa cell lysate
    Lane 3: HA750156 IP in HeLa cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 24 seconds

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