Anti-MUC1 antibody [EPR1023] is a rabbit recombinant monoclonal antibody that is used to detect MUC1 in Flow cytometry (Intra), ICC/IF, IHC-P, IP, Western blot. Suitable for Human, Mouse, Rat samples.
- Cited in over 50 publications
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Specificity confirmed with MUC1 knockout cell line validation
- Specificity and sensitivity confirmed in immunohistochemistry (IHC) with multi-tissue microarray (TMA) validation
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
IHC-P | IP | WB | ICC/IF | Flow Cyt (Intra) | |
---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Tested |
Mouse | Tested | Expected | Expected | Expected | Expected |
Rat | Tested | Expected | Expected | Expected | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/250 - 1/500 | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/250 - 1/500 | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Human | Dilution info 1/250 - 1/500 | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/20 | Notes For unpurified use at 1/10 - 1/100. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000 - 1/5000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000 | Notes For unpurified use at 1/100 - 1/250. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/30 | Notes For unpurified use at 1/100 - 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
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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.
CD227, PUM, MUC1, Mucin-1, MUC-1, Breast carcinoma-associated antigen DF3, Cancer antigen 15-3, Carcinoma-associated mucin, Episialin, H23AG, Krebs von den Lungen-6, PEMT, Peanut-reactive urinary mucin, Polymorphic epithelial mucin, Tumor-associated epithelial membrane antigen, Tumor-associated mucin, CA 15-3, KL-6, PEM, EMA
Anti-MUC1 antibody [EPR1023] is a rabbit recombinant monoclonal antibody that is used to detect MUC1 in Flow cytometry (Intra), ICC/IF, IHC-P, IP, Western blot. Suitable for Human, Mouse, Rat samples.
- Cited in over 50 publications
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Specificity confirmed with MUC1 knockout cell line validation
- Specificity and sensitivity confirmed in immunohistochemistry (IHC) with multi-tissue microarray (TMA) validation
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
This antibody 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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
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.
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 Anti-GAPDH antibody [mAbcam 9484] - Loading Control 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-MUC1 antibody [EPR1023] (ab109185) at 1/1000 dilution
Lane 1: Wild-type HeLa whole cell lysate at 20 µg
Lane 2: MUC1 knockout HeLa whole cell lysate at 20 µg
Predicted band size: 122 kDa
Observed band size: 17-24 kDa
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-MUC1 antibody [EPR1023] (ab109185) at 1/1000 dilution
All lanes: Colon cancer at 10 µg
All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 122 kDa
Observed band size: 24 kDa
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.
All lanes: Immunoprecipitation - Anti-MUC1 antibody [EPR1023] (ab109185)
Predicted band size: 122 kDa
Observed band size: 18-25 kDa
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.
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.
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.
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-MUC1 antibody [EPR1023] (ab109185) at 1/5000 dilution
All lanes: T47D cell lysate at 10 µg
All lanes: Secondary ab: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 122 kDa
Observed band size: 18-25 kDa
ab109185 (unpurified) showing positive staining in Breast ductal infiltrating carcinoma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ab109185 (unpurified) showing positive staining in Normal stomach tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Overlay histogram showing MCF7 cells fixed in 2% PFA andstained with purified ab109185 at a dilution of 1 in 30 (pink line). The secondary antibody used wasFITC goat anti-rabbit at a dilution of 1 in 150. Rabbit monoclonal IgG was used as an isotype control.
All lanes: Western blot - Anti-MUC1 antibody [EPR1023] (ab109185) at 1/1000 dilution
Lane 1: T47-D cell lysate at 10 µg
Lane 2: Human colon cancer lysate at 10 µg
Performed under reducing conditions.
Predicted band size: 122 kDa
Observed band size: 17 kDa
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 Fluorr® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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.
We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.
Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as loading control at 1/1000000 dilution.
No expression of MUC1 was detected in normal liver (PMID:16094706).
All lanes: Western blot - Anti-MUC1 antibody [EPR1023] (ab109185) at 1/1000 dilution
Lane 1: Mouse lung tissue lysate at 20 µg
Lane 2: Rat lung tissue lysate at 20 µg
Lane 3: Mouse liver tissue lysate at 20 µg
Lane 4: Rat liver tissue lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 122 kDa
Observed band size: 17-24 kDa
Exposure time: 20s
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