PRODUCT CODE: EM1902-32

MUC1 Mouse Monoclonal Antibody [A4D9] (EM1902-32)

Applications

  • IHC-P

  • FC

REACTIVITY

  • Human

Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-MUC1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1902-32, 1/100)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-MUC1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1902-32, 1/100)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-MUC1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1902-32, 1/400)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human uterus tissue using anti-MUC1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1902-32, 1/100)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-MUC1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1902-32, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-MUC1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1902-32, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of MUC1 was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1902-32, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-MUC1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1902-32, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Applications

  • IHC-P

  • FC

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Mouse monoclonal primary

Product Name

MUC1 Mouse Monoclonal Antibody [A4D9] (EM1902-32)

Immunogen

Synthetic peptide of core peptide domain of human muc1.

Host

Mouse

Positive Control

Human breast tissue, human kidney tissue, human uterus tissue, human colon carcinoma tissue, human breast carcinoma tissue, MCF-7.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

A4D9

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

2 ug/ul

PURIFICATION

Protein G affinity purified.

MOLECULAR WEIGHT

Predicted band size 122 kDa

Isotype

IgG1

APPLICATION DILUTION

  • IHC-P:1:50-1:500

  • FC:1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

MUC1

SYNONYMS

ADMCKD antibody; ADMCKD1 antibody; Breast carcinoma associated antigen DF3 antibody; Breast carcinoma-associated antigen DF3 antibody; CA 15-3 antibody; CA15 3 antibody; CA15 3 antigen antibody; CA15-3 antibody; CA15.3 antibody; Cancer antigen 15-3 antibody; Carcinoma associated mucin antibody; Carcinoma-associated mucin antibody; CD 227 antibody; CD227 antibody; DF3 antigen antibody; EMA antibody; Episialin antibody; Epithelial Membrane Antigen antibody; H23 antigen antibody; H23AG antibody; KL 6 antibody; KL-6 antibody; KL6 antibody; Krebs von den Lungen-6 antibody; MAM 6 antibody; MAM6 antibody; MCD antibody; MCKD antibody; MCKD1 antibody; Medullary cystic kidney disease 1 (autosomal dominant) antibody; Medullary cystic kidney disease, autosomal dominant antibody; MUC 1 antibody; MUC-1 antibody; MUC-1/SEC antibody; MUC-1/X antibody; MUC1 antibody; MUC1-alpha antibody; MUC1-beta antibody; MUC1-CT antibody; MUC1-NT antibody; MUC1/ZD antibody; MUC1_HUMAN antibody; Mucin 1 antibody; Mucin 1 cell surface associated antibody; Mucin 1 transmembrane antibody; Mucin 1, cell surface associated antibody; Mucin-1 subunit beta antibody; Peanut reactive urinary mucin antibody; Peanut-reactive urinary mucin antibody; PEM antibody; PEMT antibody; Polymorphic epithelial mucin antibody; PUM antibody; Tumor associated epithelial membrane antigen antibody; Tumor associated epithelial mucin antibody; Tumor associated mucin antibody; Tumor-associated epithelial membrane antigen antibody; Tumor-associated mucin antibody

TISSUE SPECIFICITY

Expressed on the apical surface of epithelial cells, especially of airway passages, breast and uterus. Also expressed in activated and unactivated T-cells. Overexpressed in epithelial tumors, such as breast or ovarian cancer and also in non-epithelial tumor cells. Isoform Y is expressed in tumor cells only.

DEVELOPMENTAL STAGE

During fetal development, expressed at low levels in the colonic epithelium from 13 weeks of gestation.

POST-TRANSLATIONAL MODIFICATION

Highly glycosylated (N- and O-linked carbohydrates and sialic acid). O-glycosylated to a varying degree on serine and threonine residues within each tandem repeat, ranging from mono- to penta-glycosylation. The average density ranges from about 50% in human milk to over 90% in T47D breast cancer cells. Further sialylation occurs during recycling. Membrane-shed glycoproteins from kidney and breast cancer cells have preferentially sialyated core 1 structures, while secreted forms from the same tissues display mainly core 2 structures. The O-glycosylated content is overlapping in both these tissues with terminal fucose and galactose, 2- and 3-linked galactose, 3- and 3,6-linked GalNAc-ol and 4-linked GlcNAc predominating. Differentially O-glycosylated in breast carcinomas with 3,4-linked GlcNAc. N-glycosylation consists of high-mannose, acidic complex-type and hybrid glycans in the secreted form MUC1/SEC, and neutral complex-type in the transmembrane form, MUC1/TM.; Proteolytic cleavage in the SEA domain occurs in the endoplasmic reticulum by an autoproteolytic mechanism and requires the full-length SEA domain as well as requiring a Ser, Thr or Cys residue at the P + 1 site. Cleavage at this site also occurs on isoform MUC1/X but not on isoform MUC1/Y. Ectodomain shedding is mediated by ADAM17.; Dual palmitoylation on cysteine residues in the CQC motif is required for recycling from endosomes back to the plasma membrane.; Phosphorylated on tyrosines and serine residues in the C-terminal. Phosphorylation on tyrosines in the C-terminal increases the nuclear location of MUC1 and beta-catenin. Phosphorylation by PKC delta induces binding of MUC1 to beta-catenin/CTNNB1 and thus decreases the formation of the beta-catenin/E-cadherin complex. Src-mediated phosphorylation inhibits interaction with GSK3B. Src- and EGFR-mediated phosphorylation on Tyr-1229 increases binding to beta-catenin/CTNNB1. GSK3B-mediated phosphorylation on Ser-1227 decreases this interaction but restores the formation of the beta-cadherin/E-cadherin complex. On T-cell receptor activation, phosphorylated by LCK. PDGFR-mediated phosphorylation increases nuclear colocalization of MUC1CT and CTNNB1.; The N-terminal sequence has been shown to begin at position 24 or 28.

SUBCELLULAR LOCATION

Apical cell membrane. Secreted. Nucleus, Cell membrane, Cytoplasm.

FUNCTION

This gene encodes a membrane-bound protein that is a member of the mucin family. Mucins are O-glycosylated proteins that play an essential role in forming protective mucous barriers on epithelial surfaces. These proteins also play a role in intracellular signaling. This protein is expressed on the apical surface of epithelial cells that line the mucosal surfaces of many different tissues including lung, breast stomach and pancreas. This protein is proteolytically cleaved into alpha and beta subunits that form a heterodimeric complex. The N-terminal alpha subunit functions in cell-adhesion and the C-terminal beta subunit is involved in cell signaling. Overexpression, aberrant intracellular localization, and changes in glycosylation of this protein have been associated with carcinomas. This gene is known to contain a highly polymorphic variable number tandem repeats (VNTR) domain. Alternate splicing results in multiple transcript variants.