MA1-90954

antibody from Invitrogen Antibodies

Targeting: MUC1 ADMCKD, ADMCKD1, CD227, MCD, MCKD, MCKD1, PEM, PUM

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Recommended by provider)

Antibody Data

Product number

MA1-90954

Provider

Invitrogen Antibodies

Product name

MUC1 Monoclonal Antibody (JZ201)

Antibody type

Monoclonal

Antigen

Other

Description

Recommended positive controls: Breast CA, colon CA.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

JZ201

Vial size

500 µL

Concentration

0.2 mg/mL

Storage

4° C, do not freeze

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-MUC1 Monoclonal Antibody (JZ201)(Product # MA1-90954) and a 160-460kdakDa band corresponding to MUC1 was observed across cell lines tested. Membrane enriched extracts (50 µg lysate) of T-47D (Lane 1), DU 145 (Lane 2), HCT 116 (Lane 3), IMR-32 (Lane 4) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:20000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556).A streak like pattern was observed in the positive cell lines

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of MUC1 was performed using 70% confluent log phase T-47D cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with MUC1 Monoclonal Antibody (JZ201) (Product # MA1-90954) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Plasma membrane and cytoplasm localization. Panel e represents HCT116. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.