Anti-MUC1 antibody [EPR1023] (ab109185)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1023] to MUC1
- Suitable for: ICC/IF, WB, IP, IHC-P, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-MUC1 antibody [EPR1023]
See all MUC1 primary antibodies -
Description
Rabbit monoclonal [EPR1023] to MUC1 -
Host species
Rabbit -
Specificity
ab109185 detects the 17 kDa carboxy-terminal subunit (subunit beta). Depending on the N-glycosylation extent, the size of this subunit is estimated to be between 17 kDa (without N-glycosylation) and 23-25 kDa -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF HumanIHC-P RatHumanIP HumanWB MouseHuman -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- T47-D cell lysate, colon cancer lysate, Human breast carcinoma tissue, Human normal breast tissue.
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General notes
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.
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 8.90 x 10 -12 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 59% PBS, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR1023 -
Isotype
IgG -
Research areas
Images
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Immunohistochemical staining of paraffin embedded human endometrium with purified ab109185 at a working dilution of 1 in 500. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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All lanes : Anti-MUC1 antibody [EPR1023] (ab109185) at 1/1000 dilution
Lane 1 : Wild-type HeLa whole cell lysate
Lane 2 : MUC1 knockout HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 17 kDa
Observed band size: 17-24 kDa why is the actual band size different from the predicted?Lanes 1 - 2: Merged signal (red and green). Green - ab109185 observed at 24 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab109185 was shown to specifically react with MUC1 in wild-type HeLa cells as signal was lost in MUC1 knockout cells. Wild-type and MUC1 knockout samples were subjected to SDS-PAGE. Ab109185 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Anti-MUC1 antibody [EPR1023] (ab109185) at 1/1000 dilution (Purified antibody) + Colon cancer at 10 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 17 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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Immunohistochemical staining of paraffin embedded mouse lung with purified ab109185 at a working dilution of 1 in 500. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Immunohistochemical staining of paraffin embedded rat kidney with purified ab109185 at a working dilution of 1 in 500. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Immunofluorescence staining of A431 cells with purified ab109185 at a working dilution of 1 in 1000, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 555 goat anti rabbit, used at a dilution of 1 in 400. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, purified ab109185 was used at a dilution of 1/200 followed by an Alexa Fluor® 555 goat anti-mouse antibody at a dilution of 1/500.
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Anti-MUC1 antibody [EPR1023] (ab109185) at 1/5000 dilution (Purified) + T47D cell lysate at 10 µg
Secondary
Secondary ab: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 17 kDa
Observed band size: 18-25 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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ab109185 (unpurified) showing positive staining in Breast ductal infiltrating carcinoma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ab109185 (unpurified) showing positive staining in Normal stomach tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ab109185 (purified) at 1/20 immunoprecipitating MUC1 in T47-D (Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Overlay histogram showing MCF7 cells fixed in 2% PFA and stained with purified ab109185 at a dilution of 1 in 30 (pink line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 150. Rabbit monoclonal IgG was used as an isotype control.
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All lanes : Anti-MUC1 antibody [EPR1023] (ab109185) at 1/1000 dilution (Unpurified)
Lane 1 : T47-D cell lysate
Lane 2 : Human colon cancer lysate
Lysates/proteins at 10 µg per lane.
Performed under reducing conditions.
Predicted band size: 17 kDa
Additional bands at: 17 kDa (possible cleavage fragment)
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Overlay histogram showing MCF7 cells stained with unpurified ab109185 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109185, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in MCF7 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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Equilibrium disassociation constant (KD) of unpurified ab109185.
Learn more about KD
Click here to learn more about KD -