WB
Western blot analysis of Calreticulin on different lysates with Rabbit anti-Calreticulin antibody at 1/5,000 dilution. Lane 1: HeLa cell lysate, Lane 2: HepG2 cell lysate, Lane 3: HL-60 cell lysate, Lane 4: NIH/3T3 cell lysate, Lane 5: C2C12 cell lysate, Lane 6: Mouse liver tissue lysate, Lane 7: Rat liver tissue lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 20 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 at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Calreticulin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/50) 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.ICC/IF
Immunocytochemistry analysis of HeLa cells labeling Calreticulin with Rabbit anti-Calreticulin antibody at 1/2,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 Rabbit anti-Calreticulin antibody at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of HeLa cells labeling Calreticulin. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
Calreticulin was immunoprecipitated in 0.2mg HeLa cell lysate with Rabbit anti-Calreticulin antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-Calreticulin antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody 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 Rabbit anti-Calreticulin antibody in HeLa cell lysate Lane 3: Rabbit anti-Calreticulin antibody IP in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 24 seconds| Product Name | Calreticulin Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within Human Calreticulin aa 40-89 / 417. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:5000-1:50000 IHC:1:50 ICC:1:2000 FC:1:2000 IP:Use at an assay dependent concentration. |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | CALR |
|---|---|
| Gene Synonyms | RO CRT SSA CALR1 cC1qR HEL-S-99n |
| Gene Full Name | calreticulin |
| Gene Summary | Calreticulin is a highly conserved chaperone protein which resides primarily in the endoplasmic reticulum, and is involved in a variety of cellular processes, among them, cell adhesion. Additionally, it functions in protein folding quality control and calcium homeostasis. Calreticulin is also found in the nucleus, suggesting that it may have a role in transcription regulation. Systemic lupus erythematosus is associated with increased autoantibody titers against calreticulin. Recurrent mutations in calreticulin have been linked to various neoplasms, including the myeloproliferative type.[provided by RefSeq, May 2020] |
| Alternative Names | Autoantigen RO antibody CALR antibody CALR protein antibody CALR_HUMAN antibody Calregulin antibody Calreticulin antibody cC1qR antibody CRP55 antibody CRT antibody CRTC antibody Autoantigen RO antibody CALR antibody CALR protein antibody CALR_HUMAN antibody Calregulin antibody Calreticulin antibody cC1qR antibody CRP55 antibody CRT antibody CRTC antibody Endoplasmic reticulum resident protein 60 antibody Epididymis secretory sperm binding protein Li 99n antibody ERp60 antibody FLJ26680 antibody grp60 antibody HACBP antibody HEL S 99n antibody RO antibody Sicca syndrome antigen A (autoantigen Ro calreticulin) antibody Sicca syndrome antigen A antibody SSA antibody |
| Molecular Weight(MW) | 48kDa(Observed band size: 55kDa) |
| Cellular Localization | Endoplasmic reticulum lumen, Cytoplasm, Secreted, Cell surface, Sarcoplasmic reticulum lumen, nucleus. |
WB
Western blot analysis of Calreticulin on different lysates with Rabbit anti-Calreticulin antibody at 1/5,000 dilution. Lane 1: HeLa cell lysate, Lane 2: HepG2 cell lysate, Lane 3: HL-60 cell lysate, Lane 4: NIH/3T3 cell lysate, Lane 5: C2C12 cell lysate, Lane 6: Mouse liver tissue lysate, Lane 7: Rat liver tissue lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 20 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 at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Calreticulin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/50) 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.
ICC/IF
Immunocytochemistry analysis of HeLa cells labeling Calreticulin with Rabbit anti-Calreticulin antibody at 1/2,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 Rabbit anti-Calreticulin antibody at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HeLa cells labeling Calreticulin. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
Calreticulin was immunoprecipitated in 0.2mg HeLa cell lysate with Rabbit anti-Calreticulin antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-Calreticulin antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody 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 Rabbit anti-Calreticulin antibody in HeLa cell lysate Lane 3: Rabbit anti-Calreticulin antibody IP in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 24 seconds| Application Notes | WB:1:5000-1:50000 IHC:1:50 ICC:1:2000 FC:1:2000 IP:Use at an assay dependent concentration. |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
