Calreticulin Recombinant Rabbit Monoclonal Antibody [SU37-03]
cat.: ET1608-60
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IHC-P, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | SU37-03 |
| Form: | Liquid |
| Storage condition: | 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. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 48 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within Human Calreticulin aa 40-89 / 417. |
| 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, HeLa, human liver tissue, human liver carcinoma tissue, human kidney tissue, mouse brain tissue. |
| Subcellular location: | Endoplasmic reticulum lumen, Cytoplasm, Secreted, Cell surface, Sarcoplasmic reticulum lumen, nucleus. |
| Recommended Dilutions:
WB IF-Cell IHC-P FC IP |
1:50,000 1:2,000 1:5,000 1:2,000 1-2μg/sample |
| Uniprot #: | SwissProt: P27797 Human | P14211 Mouse | P18418 Rat |
| Alternative names: | Autoantigen RO CALR CALR protein CALR_HUMAN Calregulin Calreticulin cC1qR CRP55 CRT CRTC Endoplasmic reticulum resident protein 60 Epididymis secretory sperm binding protein Li 99n ERp60 FLJ26680 grp60 HACBP HEL S 99n RO Sicca syndrome antigen A (autoantigen Ro; calreticulin) Sicca syndrome antigen A SSA |
Images
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Fig1:
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. |
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Fig2:
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. |
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Fig3:
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. |
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Fig4:
Immunocytochemistry analysis of HeLa cells labeling Calreticulin with Rabbit anti-Calreticulin antibody (ET1608-60) 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 (ET1608-60) at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig5:
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). |
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Fig6:
Calreticulin was immunoprecipitated in 0.2mg HeLa cell lysate with ET1608-60 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1608-60 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 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 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.