MA5-32131

CALR antibody from Invitrogen Antibodies

cC1qR, CRT, FLJ26680, RO, SSA

Western blot: Supportive data in Antibodypedia

Immunohistochemistry: Supportive data in Antibodypedia

Flow cytometry: Supportive data in Antibodypedia

Other assay: Supportive data in Antibodypedia

Provider product page for MA5-32131

Antibody data

Antibody Data
References [1]
Comments [0]
Validations
Western blot [6]
Immunohistochemistry [12]
Flow cytometry [2]
Other assay [2]

Submit

Product number: MA5-32131
Provider: Invitrogen Antibodies
Product name: Calreticulin Recombinant Rabbit Monoclonal Antibody (SU37-03)
Provider product page: Invitrogen Antibodies - MA5-32131
Antibody type: Monoclonal
Antigen: Synthetic peptide
Description: Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Antibody clone number: SU37-03
Vial size: 100 µL
Concentration: 1 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Calreticulin in different lysates using a Monoclonal antibody (Product #MA5-32131) at a dilution of 1:1,000. Positive control: Lane 1: SH-SY-5Y, Lane 2: HL-60.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Calreticulin Recombinant Rabbit Monoclonal Antibody (SU37-03) (Product # MA5-32131) and a ~55kDa band corresponding to Calreticulin was observed across all the cell lines tested. Whole cell extracts (30 µg lysate) of HL-60 (Lane 1), LNCaP (Lane 2), HeLa (Lane 3), MCF-7 (Lane 4), Mouse Liver (Lane 5) and Rat Liver (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Calreticulin in different lysates using a Monoclonal antibody (Product #MA5-32131) at a dilution of 1:1,000. Positive control: Lane 1: SH-SY-5Y, Lane 2: HL-60.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Calreticulin Recombinant Rabbit Monoclonal Antibody (SU37-03) (Product # MA5-32131) and a ~55kDa band corresponding to Calreticulin was observed across all the cell lines tested. Whole cell extracts (30 µg lysate) of HL-60 (Lane 1), LNCaP (Lane 2), HeLa (Lane 3), MCF-7 (Lane 4), Mouse Liver (Lane 5) and Rat Liver (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of Calreticulin was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR1007602_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of Calreticulin was performed by loading 30 µg of HeLa wild type (Lane 1), HeLa Cas9 (Lane 2) and HeLa Calreticulin KO (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Calreticulin Recombinant Rabbit Monoclonal Antibody (SU37-03) (Product # MA5-32131, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:10,000 dilution) using the iBright™ FL1500 (Product # A44115). Chemiluminescent detection was performed usingSuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to Calreticulin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Calreticulin Recombinant Rabbit Monoclonal Antibody (SU37-03) (Product # MA5-32131) and a ~55kDa band corresponding to Calreticulin was observed across all the cell lines tested. Whole cell extracts (30 µg lysate) of HL-60 (Lane 1), LNCaP (Lane 2), HeLa (Lane 3), MCF-7 (Lane 4), Mouse Liver (Lane 5) and Rat Liver (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemical analysis of Calreticulin in Hela cells using a Calreticulin Monoclonal antibody (Product # MA5-32131) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemical analysis of Calreticulin in MCF-7 cells using a Calreticulin Monoclonal antibody (Product # MA5-32131) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemical analysis of Calreticulin in NIH/3T3 cells using a Calreticulin Monoclonal antibody (Product # MA5-32131) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemical analysis of Calreticulin in L6 cells using a Calreticulin Monoclonal antibody (Product # MA5-32131) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Calreticulin of paraffin-embedded Human liver tissue using a Calreticulin Monoclonal antibody (Product #MA5-32131). Counter stained with hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Calreticulin of paraffin-embedded Human liver cancer tissue using a Calreticulin Monoclonal antibody (Product #MA5-32131). Counter stained with hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Calreticulin of paraffin-embedded Human kidney tissue using a Calreticulin Monoclonal antibody (Product #MA5-32131). Counter stained with hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Calreticulin of paraffin-embedded Mouse brain tissue using a Calreticulin Monoclonal antibody (Product #MA5-32131). Counter stained with hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Calreticulin of paraffin-embedded Human liver tissue using a Calreticulin Monoclonal antibody (Product #MA5-32131). Counter stained with hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Calreticulin of paraffin-embedded Human liver cancer tissue using a Calreticulin Monoclonal antibody (Product #MA5-32131). Counter stained with hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Calreticulin of paraffin-embedded Human kidney tissue using a Calreticulin Monoclonal antibody (Product #MA5-32131). Counter stained with hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Calreticulin of paraffin-embedded Mouse brain tissue using a Calreticulin Monoclonal antibody (Product #MA5-32131). Counter stained with hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Exosome purification and characterization. (A) Schematic describing exosome purification procedure. (B) Size distribution plot of purified exosomes, as determined by NTA. (C) Immunofluorescence image of purified exosomes, labeled with an Alexa Fluor 647-conjugated anti-CD63 monoclonal antibody, captured using the Particle Metrix PMX-220 ZetaView camera. (D) Electron micrograph of negative-stained, purified exosomes. Bar, 100 nm. (E) Immunoblot analysis of equal proportions of 293F cell and exosome lysates using antibodies specific for the exosomal markers CD81, CD9, CD63, E-cadherin (E-cad), N-cadherin (N-cad), GRP78/BiP, calreticulin (CALR), ERGIC-3, GM130, HSP60, and HSP90. The amount and ratio of cell and exosome lysates was selected empirically to show the different enrichment of the CD81, CD9, and CD63 proteins, was kept constant in all immunoblots, by proportion equaled a 10-fold overloading of exosomes relative to cells, and by amount of protein equaled 15 mug cell lysate protein/lane and 0.15 mug exosome lysate protein/lane.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Exosome purification and characterization. (A) Schematic describing exosome purification procedure. (B) Size distribution plot of purified exosomes, as determined by NTA. (C) Immunofluorescence image of purified exosomes, labeled with an Alexa Fluor 647-conjugated anti-CD63 monoclonal antibody, captured using the Particle Metrix PMX-220 ZetaView camera. (D) Electron micrograph of negative-stained, purified exosomes. Bar, 100 nm. (E) Immunoblot analysis of equal proportions of 293F cell and exosome lysates using antibodies specific for the exosomal markers CD81, CD9, CD63, E-cadherin (E-cad), N-cadherin (N-cad), GRP78/BiP, calreticulin (CALR), ERGIC-3, GM130, HSP60, and HSP90. The amount and ratio of cell and exosome lysates was selected empirically to show the different enrichment of the CD81, CD9, and CD63 proteins, was kept constant in all immunoblots, by proportion equaled a 10-fold overloading of exosomes relative to cells, and by amount of protein equaled 15 mug cell lysate protein/lane and 0.15 mug exosome lysate protein/lane.