Antibodies, Proteins, Kits and Reagents for Life Science

345 091 ₸ Product size

100 µl 345 091 ₸ 10 µl 140 716 ₸

Order now and get it on Tuesday March 02, 2021

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
Suitable for: WB, Flow Cyt, IP, ICC/IF
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-MUC1 antibody [EP1024Y]
See all MUC1 primary antibodies
Description

Rabbit monoclonal [EP1024Y] to MUC1
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).

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide corresponding to Human MUC1 aa 1-100 (N terminal).
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.
General notes

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 RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Primary antibody notes

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.
Clonality

Monoclonal
Clone number

EP1024Y
Isotype

IgG
Research areas

Western blot - Anti-MUC1 antibody [EP1024Y] (ab45167)

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?

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.
Immunocytochemistry/ Immunofluorescence - Anti-MUC1 antibody [EP1024Y] (ab45167)

Immunocytochemistry/Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma cell line) cells labeling purified MUC1 with ab45167 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei counterstained with DAPI (blue).

Confocal image showing membranous staining on MCF7 cell line.

Negative control: HCT-116 (PMID: 14998492).
Flow Cytometry - Anti-MUC1 antibody [EP1024Y] (ab45167)

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).
Western blot - Anti-MUC1 antibody [EP1024Y] (ab45167)

All lanes : Anti-MUC1 antibody [EP1024Y] (ab45167) at 1/1000 dilution (purified)

Lane 1 : Human colon cancer lysate
Lane 2 : Rat liver lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Observed band size: 27 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM /TBST.
Immunoprecipitation - Anti-MUC1 antibody [EP1024Y] (ab45167)

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 Detection Reagent (HRP) was used for detection (1/1000).

Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM /TBST.
Flow Cytometry - Anti-MUC1 antibody [EP1024Y] (ab45167)

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.
Western blot - Anti-MUC1 antibody [EP1024Y] (ab45167)

Anti-MUC1 antibody [EP1024Y] (ab45167) at 1/5000 dilution (purified) + Mouse liver lysate at 20 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Observed band size: 27 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-MUC1 antibody [EP1024Y] (ab45167)

Anti-MUC1 antibody [EP1024Y] (ab45167) at 1/2000 dilution (unpurified) + T47D cell lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Observed band size: 27 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking and diluting buffer: 5% NFDM/TBST.
Anti-MUC1 antibody [EP1024Y] (ab45167)

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