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Calreticulin Antibody

Novus Biologicals, part of Bio-Techne | Catalog # NB600-103

Novus Biologicals, part of Bio-Techne
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NB600-103
NB600-103SS

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Knockdown Validated, Knockout Validated, Simple Western, Western Blot

Cited:

Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

This product is unpurified. The exact concentration of antibody is not quantifiable.

Product Specifications

Immunogen

Calreticulin Antibody was developed against a full-length protein to mouse Calreticulin [UniProt# P14211]

Localization

Calreticulin staining is localized to the ER in most cell types and under most conditions.

Marker

Endoplasmic Reticulum Marker

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

48 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for Calreticulin Antibody

Knockout Validated: Calreticulin Antibody [NB600-103]

Knockout Validated: Calreticulin Antibody [NB600-103]

Knockout Validated: Calreticulin Antibody [NB600-103] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Calreticulin knockout (KO) HeLa cell line. PVDF membrane was probed with 1:2500 of Rabbit Anti-Human Calreticulin Polyclonal Antibody (Catalog # NB600-103) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog #HAF008). Specific band was detected for Calreticulin at approximately 55 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. This experiment was conducted under reducing conditions.
Simple Western: Calreticulin Antibody [NB600-103]

Simple Western: Calreticulin Antibody [NB600-103]

Simple Western: Calreticulin Antibody [NB600-103] - Lane view shows a specific band for Calreticulin in 0.2 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.
Immunocytochemistry/ Immunofluorescence: Calreticulin Antibody [NB600-103]

Immunocytochemistry/ Immunofluorescence: Calreticulin Antibody [NB600-103]

Immunocytochemistry/Immunofluorescence: Calreticulin Antibody [NB600-103] - IF Confocal analysis of 3T3 cells using Calreticulin antibody (NB600-103, 1:20). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).

Applications for Calreticulin Antibody

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:20-1:100. Use reported by customer review

Immunohistochemistry

1:20

Immunohistochemistry-Frozen

reported in scientific literature (PMID 26643212)

Immunohistochemistry-Paraffin

1:20

Knockdown Validated

1:2500

Simple Western

1:500

Western Blot

1:1000-1:10000
Application Notes
A band at approx. 55 kDa is seen in Western Blot. Note that Calreticulin has several reported isoforms that range from 48-62 kDa.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 3 reviews rated 4.7 using NB600-103 in the following applications:

Formulation, Preparation, and Storage

Purification

Unpurified

Formulation

Whole antisera

Preservative

0.1% Sodium Azide

Concentration

This product is unpurified. The exact concentration of antibody is not quantifiable.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Calreticulin

Calreticulin, also called CALR, is an endoplasmic reticulum (ER) protein that functions as a calcium (Ca2+) buffering molecular chaperone (1-3). Together with calnexin and ERp57, calreticulin has a critical role in protein folding and quality control of newly synthesized glycoproteins (1-3). Calreticulin has many diverse functions beyond Ca2+ binding, including cell adhesion, lectin binding, apoptosis, nuclear transport, antigen presentation, and immune response (1-3). Specifically, calreticulin is part of the peptide-loading complex that resides on the ER membrane and is responsible for antigen loading onto MHC class I molecules (1-3). Calreticulin protein has a theoretical molecular weight of 46 kDa and is 417 amino acids (aa) in length (1-3). The protein has three main domains: the N-terminal lectin-like domain, the proline-rich P-domain, and the acidic C-domain which is followed by an ER-retention KDEL signal sequence (1-3).

Given its role in multiple biological processes, it makes sense that calreticulin is implicated in both healthy and disease states. Studies have found that calreticulin mutations were present in patients with myeloproliferative neoplasms (MFN) and essential thrombocythaemia (ET) (4). Calreticulin expression is typically upregulated in most cancer lines, however it is downregulated in some tissues including cervical carcinomas, prostate cancer, and human colon adenocarcinoma (3). High expression of calreticulin on cancer cells is related to tumor cell phagocytosis and is often correlated with, and counteracted by, elevated CD47 expression to prevent cancer cell phagocytosis (3). Calreticulin mutations can serve as a major diagnostic biomarker for MFN and ET and additionally the calreticulin gene may be a potential target for cancer therapeutics (3,4).

References

1. Michalak, M., Groenendyk, J., Szabo, E., Gold, L. I., & Opas, M. (2009). Calreticulin, a multi-process calcium-buffering chaperone of the endoplasmic reticulum. The Biochemical Journal. https://doi.org/10.1042/BJ20081847

2. Fucikova, J., Spisek, R., Kroemer, G., & Galluzzi, L. (2021). Calreticulin and cancer. Cell Research. https://doi.org/10.1038/s41422-020-0383-9

3. Sun, J., Mu, H., Dai, K., & Yi, L. (2017). Calreticulin: a potential anti-cancer therapeutic target. Die Pharmazie. https://doi.org/10.1691/ph.2017.7031

4. Prins, D., Gonzalez Arias, C., Klampfl, T., Grinfeld, J., & Green, A. R. (2020). Mutant Calreticulin in the Myeloproliferative Neoplasms. HemaSphere. https://doi.org/10.1097/HS9.0000000000000333

Alternate Names

Autoantigen Ro, CALR, Calregulin, cC1qR, CRT, SSA, Vasostatin

Entrez Gene IDs

811 (Human)

Gene Symbol

CALR

UniProt

Additional Calreticulin Products

Product Documents for Calreticulin Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for Calreticulin Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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