PRODUCT CODE: ET1611-14

Recombinant MUC1 Monoclonal Antibody (ET1611-14)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

Western blot analysis of MUC1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1611-14, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
 Positive control: <br />
 Lane 1: Hela cell lysate<br />
 Lane 2: MCF-7 cell lysate
  • Western blot analysis of MUC1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1611-14, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
 Positive control: <br />
 Lane 1: Hela cell lysate<br />
 Lane 2: MCF-7 cell lysate
  • ICC staining of MUC1 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1611-14, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of MUC1 in B16F1 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1611-14, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human kidney 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 (ET1611-14, 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 (ET1611-14, 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-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of MUC1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1611-14, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Hela cell lysate
Lane 2: MCF-7 cell lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant MUC1 Monoclonal Antibody (ET1611-14)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

Hela cell lysate, MCF-7 cell lysate, Hela, B16F1, human kidney tissue, MCF-7.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SN06-80

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

23 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:2,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • 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; 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; 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 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, Cell membrane, Cytoplasm, Nucleus.

FUNCTION

The mucins are a family of highly glycosylated, secreted proteins with a basic structure consisting of a variable number of tandem repeats (VNTRs) encoded by 60 base pairs (Mucin 1), 69 base pairs (Mucin 2) and 51 base pairs (Mucin 3). The number of repeats is highly polymorphic and varies among different alleles. Mucin 1 proteins are expressed as type I membrane proteins in addition to secreted forms. Mucin 1 is aberrantly expressed in epithelial tumors including breast carcinomas. Mucin 2 coats the epithelia of the intestines and airways and is associated with colonic tumors. Mucin 3 is a major com-ponent of various mucus gels and is broadly expressed in normal and tumor cells.