MUC1 Recombinant Rabbit Monoclonal Antibody [SN06-80]
cat.: ET1611-14
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: SN06-80
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 122 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human MUC1 aa 1,206-1,255 / 1,255.
Positive control: HeLa cell lysate, T-47D cell lysate, mouse lung tissue lysate, rat lung tissue lysate, HeLa, B16F1, human lung carcinoma, human endometrial carcinoma, human kidney tissue.
Subcellular location: Apical cell membrane, Secreted, Cell membrane, Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
1:2,000
1:50-1:500
1:50-1:200
1:50-1:1,000
1:1,000
Use at an assay dependent concentration.
Uniprot #: SwissProt: P15941 Human | Q02496 Mouse
Entrez Gene: 24571 Rat
Alternative names: ADMCKD ADMCKD1 Breast carcinoma associated antigen DF3 Breast carcinoma-associated antigen DF3 CA 15-3 CA15 3 CA15 3 antigen CA15.3 Cancer antigen 15-3 Carcinoma associated mucin Carcinoma-associated mucin CD 227 CD227 DF3 antigen EMA Episialin H23 antigen H23AG KL 6 KL-6 KL6 Krebs von den Lungen-6 MAM 6 MAM6 MCD MCKD MCKD1 Medullary cystic kidney disease 1 (autosomal dominant) Medullary cystic kidney disease, autosomal dominant MUC 1 MUC-1 MUC-1/SEC MUC-1/X MUC1 MUC1-alpha MUC1-beta MUC1-CT MUC1-NT MUC1/ZD MUC1_HUMAN Mucin 1 Mucin 1 transmembrane Mucin 1, cell surface associated Mucin-1 subunit beta Peanut reactive urinary mucin Peanut-reactive urinary mucin PEM PEMT Polymorphic epithelial mucin PUM Tumor associated epithelial membrane antigen Tumor associated epithelial mucin Tumor associated mucin Tumor-associated epithelial membrane antigen Tumor-associated mucin
Images
ET1611-14_1.jpg Fig1: Western blot analysis of MUC1 on different lysates with Rabbit anti-MUC1 antibody (ET1611-14) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: HCT 116 cell lysate (negative) (20 µg/Lane)
Lane 3: T-47D cell lysate (20 µg/Lane)
Lane 4: Mouse lung tissue lysate (40 µg/Lane)
Lane 5: Rat lung tissue lysate (40 µg/Lane)

Predicted band size: 122 kDa
Observed band size: 17~24 kDa

Exposure time: 3 minutes 10 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-14) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1611-14_2.jpg Fig2: Western blot analysis of MUC1 on different lysates with Rabbit anti-MUC1 antibody (ET1611-14) at 1/2,000 dilution.

Lane 1: Hela-si NT cell lysate
Lane 2: Hela-si MUC1#1 cell lysate
Lane 3: Hela-si MUC1#2 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 122 kDa
Observed band size: 17~24 kDa

Exposure time: 10 seconds;

4-20% SDS-PAGE gel.

ET1611-14 was shown to specifically react with MUC1 in Hela-si NT cells. Weakened bands were observed when Hela-si MUC1 sample were tested. Hela-si NT and Hela-si MUC1 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1611-14, 1/2,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.
ET1611-14_3.jpg Fig3: Immunocytochemistry analysis of HeLa (positive) and HCT 116 (negative) labeling MUC1 with Rabbit anti-MUC1 antibody (ET1611-14) at 1/500 dilution and competitor's antibody at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MUC1 antibody (ET1611-14) at 1/500 dilution and competitor's antibody at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1611-14_4.jpg Fig4: ICC staining of MUC1 in B16F1 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1611-14, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1611-14_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue with Rabbit anti-MUC1 antibody (ET1611-14) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-14) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1611-14_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human endometrial carcinoma tissue with Rabbit anti-MUC1 antibody (ET1611-14) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-14) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1611-14_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-MUC1 antibody (ET1611-14) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-14) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1611-14_8.jpg Fig8: Flow cytometric analysis of HeLa cells labeling MUC1.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1611-14, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET1611-14_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-MUC1 antibody (ET1611-14) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-14) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1611-14_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-MUC1 antibody (ET1611-14) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-14) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1611-14_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-MUC1 antibody (ET1611-14) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-14) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.