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Calreticulin Antibody (1G6A7) [Alexa Fluor® 488]

Novus Biologicals, part of Bio-Techne | Catalog # NBP1-47518AF488

Novus Biologicals, part of Bio-Techne

Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human

Applications

Validated:

ELISA, Flow (Intracellular), Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, In vitro assay, Western Blot

Cited:

Flow Cytometry

Label

Alexa Fluor 488 (Excitation = 488 nm, Emission = 515-545 nm)

Antibody Source

Monoclonal Mouse IgG2A Clone # 1G6A7

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Product Specifications

Immunogen

Calreticulin Antibody (1G6A7) was developed against a synthetic peptide corresponding to the C-terminus (EEEDVPGQAKDELC) of human Calreticulin, conjugated to KLH. [UniProt# P27797]

Marker

Endoplasmic Reticulum Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Scientific Data Images for Calreticulin Antibody (1G6A7) [Alexa Fluor® 488]

Immunocytochemistry/Immunofluorescence: Calreticulin Antibody (1G6A7) [Alexa Fluor 488] [NBP1-47518AF488] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton-X100. The cells were incubated with anti-Calreticulin (1G6A7) conjugated to Alexa Fluor 488 [NBP1-47518AF488] at 10ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Flow Cytometry: Calreticulin Antibody (1G6A7) [Alexa Fluor® 488] [NBP1-47518AF488] - An intracellular stain was performed on HepG2 cells with Calreticulin Antibody (1G6A7) NBP1-47518AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.
Immunocytochemistry/Immunofluorescence: Calreticulin Antibody (1G6A7) [Alexa Fluor 488] [NBP1-47518AF488] - The Calreticulin antibody was tested in HeLa cells at a 1:250 dilution against Dylight 488 (Green). Actin was counterstained against Phalloidin 568 (Red) and cells were mounted in DAPI Flouromount (Blue).

Applications for Calreticulin Antibody (1G6A7) [Alexa Fluor® 488]

Application
Recommended Usage

ELISA

Optimal dilutions of this antibody should be experimentally determined.

Flow (Intracellular)

Optimal dilutions of this antibody should be experimentally determined.

Flow Cytometry

Optimal dilutions of this antibody should be experimentally determined.

Immunocytochemistry/ Immunofluorescence

Optimal dilutions of this antibody should be experimentally determined.

Immunohistochemistry

Optimal dilutions of this antibody should be experimentally determined.

Immunohistochemistry-Paraffin

Optimal dilutions of this antibody should be experimentally determined.

In vitro assay

Optimal dilutions of this antibody should be experimentally determined.

Western Blot

Optimal dilutions of this antibody should be experimentally determined.
Application Notes
Optimal dilution of this antibody should be experimentally determined.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Ammonium sulfate precipitation

Formulation

50mM Sodium Borate

Preservative

0.05% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

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

Stability & Storage

Store at 4C in the dark.

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

Gene Symbol

CALR

Additional Calreticulin Products

Product Documents for Calreticulin Antibody (1G6A7) [Alexa Fluor® 488]

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 (1G6A7) [Alexa Fluor® 488]

Alexa Fluor (R) products are provided under an intellectual property license from Life Technologies Corporation. The purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: (i) in manufacturing; (ii) to provide a service, information, or data in return for payment; (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com. This conjugate is made on demand. Actual recovery may vary from the stated volume of this product. The volume will be greater than or equal to the unit size stated on the datasheet.

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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