Anti-MUC1 antibody [EPR1023] - Low endotoxin, Azide free (ab194978)

Rabbit Recombinant Monoclonal MUC1 antibody. Carrier free. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Mouse, Rat, Human samples. Cited in 2 publications.

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC1 antibody [EPR1023] - Low endotoxin, Azide free (AB194978), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	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

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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Target data

See full target information MUC1

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.

Alternative names

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

Recommended products

Rabbit Recombinant Monoclonal MUC1 antibody. Carrier free. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Mouse, Rat, Human samples. Cited in 2 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR1023
Purification technique: Affinity purification Protein A
Specificity: 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
Dissociation constant: 8.9 x 10^-12 M
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

ab194978 is the carrier-free version of Anti-MUC1 antibody [EPR1023] ab109185.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

What does low endotoxin mean?
Our low endotoxin, azide-free formats have low endotoxin level (1 EU/mg, determined by the TAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

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.

12 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC1 antibody [EPR1023] - Low endotoxin, Azide free (ab194978)
Immunohistochemical staining of paraffin embedded human endometrium with purified Anti-MUC1 antibody [EPR1023] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MUC1 antibody [EPR1023] ab109185).
Western blot - Anti-MUC1 antibody [EPR1023] - Low endotoxin, Azide free (ab194978)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab194978).
Lanes 1 - 2: Merged signal (red and green). Green - Anti-MUC1 antibody [EPR1023] ab109185 observed at 24 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.
Anti-MUC1 antibody [EPR1023] 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. Anti-MUC1 antibody [EPR1023] 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] (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
Immunoprecipitation - Anti-MUC1 antibody [EPR1023] - Low endotoxin, Azide free (ab194978)
Anti-MUC1 antibody [EPR1023] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MUC1 antibody [EPR1023] ab109185).
All lanes: Immunoprecipitation - Anti-MUC1 antibody [EPR1023] (Anti-MUC1 antibody [EPR1023] ab109185)
Predicted band size: 122 kDa
Observed band size: 18-25 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC1 antibody [EPR1023] - Low endotoxin, Azide free (ab194978)
Immunohistochemical staining of paraffin embedded mouse lung with purified Anti-MUC1 antibody [EPR1023] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MUC1 antibody [EPR1023] ab109185).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC1 antibody [EPR1023] - Low endotoxin, Azide free (ab194978)
Immunohistochemical staining of paraffin embedded rat kidney with purified Anti-MUC1 antibody [EPR1023] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MUC1 antibody [EPR1023] ab109185).
Immunocytochemistry/ Immunofluorescence - Anti-MUC1 antibody [EPR1023] - Low endotoxin, Azide free (ab194978)
Immunofluorescence staining of A431 cells with purified Anti-MUC1 antibody [EPR1023] 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 Anti-MUC1 antibody [EPR1023] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MUC1 antibody [EPR1023] ab109185).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC1 antibody [EPR1023] - Low endotoxin, Azide free (ab194978)
This IHC data was generated using the same anti-MUC1 antibody clone [EPR1023] in a different buffer formulation (cat# Anti-MUC1 antibody [EPR1023] ab109185).
Anti-MUC1 antibody [EPR1023] ab109185 (unpurified) showing positive staining in Breast ductal infiltrating carcinoma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC1 antibody [EPR1023] - Low endotoxin, Azide free (ab194978)
Anti-MUC1 antibody [EPR1023] ab109185 (unpurified) showing positive staining in Normal stomach tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MUC1 antibody [EPR1023] ab109185).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cytometry (Intracellular) - Anti-MUC1 antibody [EPR1023] - Low endotoxin, Azide free (ab194978)
Overlay histogram showing MCF7 cells fixed in 2% PFA and stained with purified Anti-MUC1 antibody [EPR1023] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MUC1 antibody [EPR1023] ab109185).
Flow Cytometry (Intracellular) - Anti-MUC1 antibody [EPR1023] - Low endotoxin, Azide free (ab194978)
Overlay histogram showing MCF7 cells stained with unpurified Anti-MUC1 antibody [EPR1023] 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 (Anti-MUC1 antibody [EPR1023] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MUC1 antibody [EPR1023] ab109185).
OI-RD Scanning - Anti-MUC1 antibody [EPR1023] - Low endotoxin, Azide free (ab194978)
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.
Western blot - Anti-MUC1 antibody [EPR1023] - Low endotoxin, Azide free (ab194978)
Blocking and dilution buffer: 5% NFDM/TBST.
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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MUC1 antibody [EPR1023] ab109185).
All lanes: Western blot - Anti-MUC1 antibody [EPR1023] (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
Secondary
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