Recombinant Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (ab218998)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1024Y] to MUC1 - Low endotoxin, Azide free
- Suitable for: Flow Cyt, IP, ICC/IF, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free
See all MUC1 primary antibodies -
Description
Rabbit monoclonal [EP1024Y] to MUC1 - Low endotoxin, Azide free -
Host species
Rabbit -
Specificity
Based on the immunogen sequence, the antibody recognises several isoforms of MUC1 (Uniprot ID P15941). They are Isoform Y (28 kDa), Isoform Y-LSP (28 kDa), Isoform S2 (17 kDa) and Isoform J13 (28 kDa).
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Tested applications
Suitable for: Flow Cyt, IP, ICC/IF, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, T47D, MCF7 and A549 cell lysates; Human kidney, human breast carcinoma, human thyroid carcinoma and human colon cancer lysates; Human fetal lung lysate; Rat liver lysate and mouse liver lysate. ICC/IF: MCF7 cells. Flow Cyt: T47D and A549 cells.
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General notes
ab218998 is the carrier-free version of ab45167.
Isoform 7 of MUC1 behaves as a receptor and binds the secreted isoform 5. The binding induces the phosphorylation of the isoform 7, alters cellular morphology and initiates cell signaling. The mouse and rat recommendation is based on WB results.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1024Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab218998 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IP |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. |
Target
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Function
The alpha subunit has cell adhesive properties. Can act both as an adhesion and an anti-adhesion protein. May provide a protective layer on epithelial cells against bacterial and enzyme attack.
The beta subunit contains a C-terminal domain which is involved in cell signaling, through phosphorylations and protein-protein interactions. Modulates signaling in ERK, SRC and NF-kappa-B pathways. In activated T-cells, influences directly or indirectly the Ras/MAPK pathway. Promotes tumor progression. Regulates TP53-mediated transcription and determines cell fate in the genotoxic stress response. Binds, together with KLF4, the PE21 promoter element of TP53 and represses TP53 activity. -
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. -
Involvement in disease
MUC1/CA 15-3 is used as a serological clinical marker of breast cancer to monitor response to breast cancer treatment and disease recurrence (PubMed:20816948). Decreased levels over time may be indicative of a positive response to treatment. Conversely, increased levels may indicate disease progression. At an early stage disease, only 21% of patients exhibit high MUC1/CA 15-3 levels, that is why CA 15-3 is not a useful screening test. Most antibodies target the highly immunodominant core peptide domain of 20 amino acid (APDTRPAPGSTAPPAHGVTS) tandem repeats. Some antibodies recognize glycosylated epitopes.
Medullary cystic kidney disease 1 -
Sequence similarities
Contains 1 SEA domain. -
Developmental stage
During fetal development, expressed at low levels in the colonic epithelium from 13 weeks of gestation. -
Post-translational
modificationsHighly 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. -
Cellular localization
Secreted; Cell membrane. Cytoplasm. Nucleus. On EGF and PDGFRB stimulation, transported to the nucleus through interaction with CTNNB1, a process which is stimulated by phosphorylation. On HRG stimulation, colocalizes with JUP/gamma-catenin at the nucleus and Apical cell membrane. Exclusively located in the apical domain of the plasma membrane of highly polarized epithelial cells. After endocytosis, internalized and recycled to the cell membrane. Located to microvilli and to the tips of long filopodial protusions. - Information by UniProt
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Database links
- Entrez Gene: 4582 Human
- Entrez Gene: 17829 Mouse
- Entrez Gene: 24571 Rat
- Omim: 158340 Human
- SwissProt: P15941 Human
- SwissProt: Q02496 Mouse
- Unigene: 89603 Human
- Unigene: 16193 Mouse
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Alternative names
- ADMCKD antibody
- ADMCKD1 antibody
- Breast carcinoma associated antigen DF3 antibody
see all
Images
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All lanes : Anti-MUC1 antibody [EP1024Y] (ab45167) at 1/1000 dilution
Lane 1 : MCF7 (Human breast adenocarcinoma cell line) cell lysate
Lane 2 : T-47D (Human ductal breast epithelial tumor cell line) cell lysate
Lane 3 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 4 : MUC1 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 24 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab45167).
Lanes 1- 4: Merged signal (red and green). Green - ab45167 observed at 24 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab45167 was shown to react with MUC1 in Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) cells in western blot. Loss of signal was observed when knockout cell line ab255412 (knockout cell lysate ab263764) was used. Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) and MUC1 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab45167 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma cell line) cells labeling MUC1 with purified ab45167 at 1/500 dilution (0.2μg/ml). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, a goat anti-rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1/1000 dilution. Ab195889, an anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used as counterstain at 1/200 dilution (2.5 μg/ml). The negative control is PBS instead of the primary antibody. Nuclei counterstained with DAPI (blue).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45167).
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ab45167 (purified) at 1/20 dilution (2ug) immunoprecipitating MUC1 in Human fetal lung lysate.
Lane 1 (input): Human fetal lung lysate 10ug
Lane 2 (+): ab45167 + Human fetal lung lysate 10ug
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab45167 in Human fetal lung lysate
For western blotting, ab131366 VeriBlot for IP (HRP) was used for detection at 1/1000 dilution.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45167).
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Flow Cytometry analysis of A549 (human lung carcinoma cell line) cells labeling MUC1 with purified ab45167 at 1/20 dilution (10 ug/ml). Cells were fixed with 4% paraformaldehyde. A goat anti-rabbit IgG (Alexa Fluor® 488) was used as the secondary antibody at 1/2000 dilution. Black - Isotype control, Rabbit monoclonal IgG. Blue - unlabeled control, cells without incubation with primary antibody and secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45167).
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Flow cytometry analysis of T47D (human mammary gland ductal carcinoma) cells labelling MUC1 with unpurified ab45167 (pink) at a dilution of 1/150. Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG was used as the secondary antibody. Rabbit monoclonal IgG (ab172730) was used as the isotype control (green).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45167).
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (5)
ab218998 has been referenced in 5 publications.
- Alysandratos KD et al. Patient-specific iPSCs carrying an SFTPC mutation reveal the intrinsic alveolar epithelial dysfunction at the inception of interstitial lung disease. Cell Rep 36:109636 (2021). PubMed: 34469722
- Gorges TM et al. Circulating tumour cells escape from EpCAM-based detection due to epithelial-to-mesenchymal transition. BMC Cancer 12:178 (2012). WB . PubMed: 22591372
- Sachdeva M & Mo YY MicroRNA-145 suppresses cell invasion and metastasis by directly targeting mucin 1. Cancer Res 70:378-87 (2010). PubMed: 19996288
- Williams MA et al. Deletion of the mucin-like molecule muc1 enhances dendritic cell activation in response to toll-like receptor ligands. J Innate Immun 2:123-43 (2010). PubMed: 20375631
- Huang Y et al. Kidney-derived stromal cells modulate dendritic and T cell responses. J Am Soc Nephrol 20:831-41 (2009). PubMed: 19297559