I am interested in purchasing MUC1 protein from you and it would be a great help if you could answer a few of my questions. 1) How much of the glycoprotein is glycosylated? 2) Is it soluble in water? 3) It says on the website that the product is not active, what exactly do you mean by that?

This protein is a recombinant protein. It does not undergo glycosylation. It may not be folded in the same way as the native protein therefore it may be not act in the same way. This should only be used for WB and as a positive control in ELISA. It is soluble in water.

Could you recommend us a protocol for ELISA test in a sandwich format for MUC1 protein catalog # H00004582-Q01 using catalog # NB100-65647 as primary antibody and catalog # NBP1-61561 as secondary with a third labeled antibody? Regarding the request below with that pair of antibodies and that recombinant protein: could you help us with another primary monoclonal antibody to replace the NB100-65647 antibody with it, so we could use it in a sandwich assay with the protein H00004582-Q01 and the NBP1-61561. Thank you. Also do you know if this recombinant protein is tumoral MUC1 or normal MUC1, because they have different structures.

Unfortunately, the antibody and protein combination you have chosen will not work. As is stated in our datasheet for NB100-65647 The dominant epitope recognized by this antibody is the 12-mer GVTSAPDTRPAP. Unfortunately, this epitope is not present in the partial recombinant protein H00004582-Q01. While NB100-65647 and NBP1-61561 are suitable pairs to use for ELISA, H00004582-Q01 is not a suitable standard for use with this pair, and unfortunately it does not look like we carry a suitable standard for you. I apologize for the inconvenience. Please see our ELISA protocol. H00004582-Q01 is produced by a Taiwanese company called Abnova and we distribute it for them. It is produced in their cell-free wheat germ expression system. I would recommend using one of Abnova's matched antibody pairs for use in ELISA if you are going to use H00004582-Q01 as a standard. I would recommend using the H00004582-AP41 antibody pair.

Recombinant Human MUC1 GST (N-Term) Protein

Name: Recombinant Human MUC1 GST (N-Term) Protein
Brand: Novus Biologicals
SKU: H00004582-Q01

Novus Biologicals, part of Bio-Techne | Catalog # H00004582-Q01

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Citations (4)

Product Datasheet / COA / SDS

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Product Specifications
Scientific Data
Preparation & Storage
Background
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Specific Notices

Key Product Details

Source

Wheat germ

Tag

GST (N-Term)

Conjugate

Unconjugated

Applications

ELISA, Affinity Purification, Microarray, Western Blot

View all MUC-1 Proteins and Enzymes »

Product Specifications

Description

A recombinant protein with a N-terminal GST tag corresponding to the amino acid sequence 315-420 of Human MUC-1

Source: Wheat Germ (in vitro)

Amino Acid Sequence: NSSLEDPSTDYYQELQRDISEMFLQIYKQGGFLGLSNIKFRPGSVVVQLTLAFREGTINVHDMETQFNQYKTEAASRYNLTISDVSVSDVPFPFSAQSGAGVPGWG

Purity

>80% by SDS-PAGE and Coomassie blue staining

Predicted Molecular Mass

37.4 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.

Activity

This protein was produced in an in vitro wheat germ expression system that should preserve correct conformational folding that is necessary for biological function. While it is possible that this protein could display some level of activity, the functionality of this protein has not been explicitly measured or validated.

Protein / Peptide Type

Recombinant Protein

Scientific Data Images for Recombinant Human MUC1 GST (N-Term) Protein

Recombinant Human MUC-1 Protein [H00004582-Q01] - 12.5% SDS-PAGE Stained with Coomassie Blue.

Formulation, Preparation and Storage

H00004582-Q01

Preparation Method	in vitro wheat germ expression system
Formulation	50 mM Tris-HCI, 10 mM reduced Glutathione, pH 8.0 in the elution buffer.
Preservative	No Preservative
Concentration	Please see the vial label for concentration. If unlisted please contact technical services.
Shipping	The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage	Store at -80C. 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

Additional MUC-1 Products

Product Documents for Recombinant Human MUC1 GST (N-Term) Protein

Product Datasheets

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COA

SDS

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Product Specific Notices for Recombinant Human MUC1 GST (N-Term) Protein

This product is produced by and distributed for Abnova, a company based in Taiwan.

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

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