WB result of Calreticulin Rabbit mAb
Primary antibody: Calreticulin Rabbit mAb at 1/1000 dilution
Lane 1: A431 whole cell lysate 20 µg
Lane 2: HeLa whole cell lysate 20 µg
Lane 3: SH-SY5Y whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 48 kDa
Observed MW: 60 kDa
Calreticulin Recombinant Rabbit mAb (S-R353)
Calreticulin Recombinant Rabbit mAb (S-R353)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Synonyms | CRP55, Calregulin, Endoplasmic reticulum resident protein 60 (ERp60), HACBP, grp60, CALR, CRTC |
| Location | Cytoplasm, Secreted |
| Accession | P27797 |
| Clone Number | S-R353 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC, ICFCM |
| Reactivity | Hu |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied. |
Dilution
| application | dilution | species |
| WB | 1:1000-1:10000 | null |
| IHC-P | 1:2000 | null |
| ICC | 1:500 | null |
| ICFCM | 1:500 | null |
Background
Calreticulin is a multifunctional soluble protein that binds Ca2+ ions (a second messenger in signal transduction), rendering it inactive. The Ca2+ is bound with low affinity, but high capacity, and can be released on a signal (see inositol trisphosphate). Calreticulin is located in storage compartments associated with the endoplasmic reticulum and is considered an ER resident protein. Calreticulin also binds to misfolded proteins and prevents them from being exported from the endoplasmic reticulum to the Golgi apparatus.
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Western Blot
FC
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized A431 (Human epidermoid carcinoma epithelial cell) labelling Calreticulin antibody at 1/500 dilution (0.1 μg) / (Red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human colon. Anti-Calreticulin antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human liver. Anti-Calreticulin antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Negative control: IHC shows negative staining in paraffin-embedded human cardiac muscle. Anti-Calreticulin antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human breast cancer. Anti-Calreticulin antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human lung squamous cell carcinoma. Anti-Calreticulin antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC shows positive staining in A431 cells. Anti- Calreticulin antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (red).
