Calreticulin Recombinant Rabbit Monoclonal Antibody [SU37-03] - BSA and Azide free
Catalog# HA750156
Calreticulin Recombinant Rabbit Monoclonal Antibody [SU37-03] - BSA and Azide free
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WB
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IHC-P
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IF-Cell
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FC
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IP
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Human
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Mouse
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Rat
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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
Target 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
Verified (internally validated)
Reported in literature (not internally validated)
Predicted reactive (based on sequence homology)
Not recommended (failed internal validation)
| WB | IHC-P | FC | IF-Cell | IP | |
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| human |
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| mouse |
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| rat |
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Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at 2-8℃. Avoid freeze.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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.
Synonyms
Autoantigen RO antibody
CALR antibody
CALR protein antibody
CALR_HUMAN antibody
Calregulin antibody
Calreticulin antibody
cC1qR antibody
CRP55 antibody
CRT antibody
CRTC antibody
ExpandImages
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This data was developed using ET1608-60, the same antibody clone in a different buffer formulation.
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: ET1608-60, 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). -
This data was developed using ET1608-60, the same antibody clone in a different buffer formulation.
Western blot analysis of Calreticulin on different lysates with Rabbit anti-Calreticulin antibody (ET1608-60) 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 (ET1608-60) 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. -
☑ Knockdown (KD)
This data was developed using ET1608-60, the same antibody clone in a different buffer formulation.
Western blot analysis of Calreticulin on different lysates with Rabbit anti-Calreticulin antibody (ET1608-60) 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 (ET1608-60) 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. -
☑ Knockdown (KD)
This data was developed using ET1608-60, the same antibody clone in a different buffer formulation.
Western blot analysis of Calreticulin on different lysates with Rabbit anti-Calreticulin antibody (ET1608-60) 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 (ET1608-60) 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. -
This data was developed using ET1608-60, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Calreticulin antibody (ET1608-60) 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 (ET1608-60) 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. -
This data was developed using ET1608-60, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue with Rabbit anti-Calreticulin antibody (ET1608-60) 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 (ET1608-60) 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. -
This data was developed using ET1608-60, the same antibody clone in a different buffer formulation.
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: ET1608-60, 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). -
This data was developed using ET1608-60, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of HeLa cells labeling Calreticulin.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1608-60, 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). -
This data was developed using ET1608-60, the same antibody clone in a different buffer formulation.
Calreticulin was immunoprecipitated in 0.2mg HeLa cell lysate with ET1608-60 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1608-60 at 1/1,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) 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 ET1608-60 in HeLa cell lysate
Lane 3: ET1608-60 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"