MA1-12592

antibody from Invitrogen Antibodies

Targeting: KRT7 CK7, K2C7, K7, SCL

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Recommended by provider)

Antibody Data

Product number

MA1-12592

Provider

Invitrogen Antibodies

Product name

Cytokeratin 7 Monoclonal Antibody (C35)

Antibody type

Monoclonal

Antigen

Other

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

C35

Vial size

100 µg

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Cytokeratin 7 Monoclonal Antibody (C35) (Product # MA1-12592) and a 51 kDa band corresponding to KRT-7 was observed in positive cell lines HeLa, A549, OVCAR3 and not in negative cell lines such as A-431 and SK-OV-3 as mentioned in literature. Membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), A549 (Lane 2), OVCAR3 (Lane 3) and A-431 (Lane 4) and SK-OV-3 (Lane 5) were electrophoresed using NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) 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, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Cytokeratin 7 was performed using 70% confluent HeLa 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 1 hour at room temperature. The cells were labeled with Cytokeratin 7 Monoclonal Antibody (C35) (Product # MA1-12592) at 1:200 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 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 cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.