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MUC1 Antibody (Mc5) - Chimeric

Novus Biologicals, part of Bio-Techne | Catalog # NBP3-21451

Recombinant Monoclonal Antibody
Novus Biologicals, part of Bio-Techne

Key Product Details

Species Reactivity

Human

Applications

ELISA, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Kappa Clone # Mc5

Concentration

1 mg/ml

Product Specifications

Immunogen

Human breast epithelial mucin

Specificity

This antibody is specific for the amino acid sequence DTRPAP of MUC1

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG Kappa

Description

This chimeric rabbit antibody was made using the variable domain sequences of the original Mouse IgG2 format, for improved compatibility with existing reagents, assays and techniques.

Scientific Data Images for MUC1 Antibody (Mc5) - Chimeric

MUC1 Antibody (Mc5) - Chimeric

Immunocytochemistry/ Immunofluorescence: Rabbit MUC1 mAb (Mc5) - Chimeric [NBP3-21451] -

Immunocytochemistry/ Immunofluorescence: Rabbit MUC1 mAb (Mc5) - Chimeric [NBP3-21451] - Immunofluorescence analysis of paraformaldehyde fixed MCF7 cells permeabilized with 0.15% Triton and stained with the chimeric mouse IgG1 version of Mc5 at 10 ug/ml for1h followed by Alexa Fluor® 488 secondary antibody (2 ug/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom DAPI, merged channels and an isotype control. The isotype control was stained with an anti-unknown specificity antibody followed by Alexa Fluor® 488 secondary antibody.
MUC1 Antibody (Mc5) - Chimeric

Western Blot: Rabbit MUC1 mAb (Mc5) - Chimeric [NBP3-21451] -

Western Blot: Rabbit MUC1 mAb (Mc5) - Chimeric [NBP3-21451] - MCF7 cell lysate (35µg protein in RIPA buffer) was resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit IgG version of Mc5 at 2 µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence. The expected running size for unmodified breast epithelial mucin is 122.1kDa, but this protein is heavily glycosylated and lipidated at several positions leading to the observed running size.

Applications for MUC1 Antibody (Mc5) - Chimeric

Application
Recommended Usage

ELISA

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.

Immunoprecipitation

Optimal dilutions of this antibody should be experimentally determined.

Western Blot

Optimal dilutions of this antibody should be experimentally determined.
Application Notes
In an attempt to develop a reliable way to detect breast cancer, immunoprecipitation was preformed on NPGP from HMFG using the mouse version of this antibody bound to Sepharose 4B. Furthermore, this antibody was tested in an radioimunnology experiment on NPGP using the mouse version of this antibody (Ceriani et al, 1992; pmid:1377884). The epitope of this antibody was mapped by ELISA using small peptides - the murine version of this antibody was used (Petrakou et al, 1998 ; pmid:9422085). This antibody was used to identify cells expressing the MUC1 protein. This was accomplished by using the mouse version of this antibody on cell lysates of cell transfected with the MUC1 protein in a western blot analysis (Rubinstein et al, 2006; pmid:17145869). To compare the diagnostic usefulness of different antibodies, the mouse version of this antibody was used for immunohistochemistry on cell pellets from pleural fluid specimens (Creaney et al, 2008; pmid:18454162). While developing an antibody against the alpha/beta junction of MUC1 the mouse version of this antibody was used for western blot on the MUC1 protein (Rubinstein et al, 2008; pmid:18821582). While assesing a panel of 56 anti MUC1 antibodies for their ability to detect cancer the mouse version of this antibody was used for an immunoradiometric assay on a serum made from sera from 10 different patients with breast, ovarian or colon cancer (Norum et al, 1998; pmid:9422099). To more accurately determine the epitope of this antibody, an ELISA was preformed using the murine version of this antibody on different small pepetides from the MUC1 protein (Karsten et al, 2004; pmid:15115750).

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS

Preservative

0.02% Proclin 300

Concentration

1 mg/ml

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: MUC-1

Mucin 1 (MUC1), also known as episialin, EMA (epithelial membrane antigen), PEM (polymorphic epithelial mucin), and CA-15-3 antigen, is a membrane-bound type I transmembrane glycoprotein (1,2). MUC1 is typically expressed in the luminal or glandular epithelial cells of the gastrointestinal tract, breast, lungs, and more, and is often overexpressed in epithelial cancers, but also serves a protective role against infection and helps regulate inflammatory response (2,3). Human MUC1 is 1255 amino acids (aa) in length with a theoretical molecular weight of 122 kDa; however, depending on the amount of glycosylation can weigh between 250 - 500 kDa (2,4). Structurally, MUC1 consists of a N-terminal domain which contains a signal peptide, a variable number tandem repeat region (VNTR), and a SEA domain, as well a C-terminal domain which has the extracellular domain (ECD), transmembrane domain (TMD), and cytoplasmic tail (CT) (2,3). The VNTR is comprised of between 25 - 125 repeats of a 20 aa conserved sequence (3). MUC1 is heavily O-glycosylated in the VNTR and has moderate N-glycosylation sites following the VNTR and in the ECD (2). Glycosylation contributes to MUC1's functional properties (2). The MUC1 gene contains seven exons, giving rise to several MUC1 isoforms as a result of alternative splicing (2).

Overexpression of mucins, including MUC1, is a feature of many epithelial cancers (1,3,5,6). The presence of truncated glycan structures called tumor-associated carbohydrate antigens (TACAs) on MUC1 play a role in cancer progression and a loss of apical-basal polarity (5). Carbohydrate-binding partners called lectins are the primary binding partners of TACAs that give rise to the pro-tumor microenvironment and metastasis (5). Given this unique feature, TACAs are a potential target for cancer immunotherapies (5). There are a number of vaccines, drugs, and antibodies targeting MUC1 for treatment of a variety of cancers including breast, lung, and prostate (6). In addition to a role in cancer progression, MUC1, and specifically the CT portion, has been shown to have a positive, anti-inflammatory role in a variety of lung and airway infections (7).

References

1. Khodabakhsh, F., Merikhian, P., Eisavand, M. R., & Farahmand, L. (2021). Crosstalk between MUC1 and VEGF in angiogenesis and metastasis: a review highlighting roles of the MUC1 with an emphasis on metastatic and angiogenic signaling. Cancer cell international. https://doi.org/10.1186/s12935-021-01899-8

2. Nath, S., & Mukherjee, P. (2014). MUC1: a multifaceted oncoprotein with a key role in cancer progression. Trends in molecular medicine. https://doi.org/10.1016/j.molmed.2014.02.007

3. Dhar, P., & McAuley, J. (2019). The Role of the Cell Surface Mucin MUC1 as a Barrier to Infection and Regulator of Inflammation. Frontiers in cellular and infection microbiology. https://doi.org/10.3389/fcimb.2019.00117

4. Uniprot (P15941)

5. Beckwith, D. M., & Cudic, M. (2020). Tumor-associated O-glycans of MUC1: Carriers of the glyco-code and targets for cancer vaccine design. Seminars in immunology. https://doi.org/10.1016/j.smim.2020.101389

6. Almasmoum H. (2021). The Roles of Transmembrane Mucins Located on Chromosome 7q22.1 in Colorectal Cancer. Cancer management and research. https://doi.org/10.2147/CMAR.S299089

7. Ballester, B., Milara, J., & Cortijo, J. (2021). The role of mucin 1 in respiratory diseases. European respiratory review : an official journal of the European Respiratory Society. https://doi.org/10.1183/16000617.0149-2020

Long Name

Mucin 1, Cell Surface-associated

Alternate Names

CD227, Episialin, H23AG, KL-6, Mucin-1, PEM, PEMT

Gene Symbol

MUC1

UniProt

Additional MUC-1 Products

Product Documents for MUC1 Antibody (Mc5) - Chimeric

Certificate of Analysis

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

Product Specific Notices for MUC1 Antibody (Mc5) - Chimeric

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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