MA1-35546

antibody from Invitrogen Antibodies

Targeting: KRT18

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Recommended by provider)

Antibody Data

Product number

MA1-35546

Provider

Invitrogen Antibodies

Product name

Cytokeratin 18 Monoclonal Antibody (M9)

Antibody type

Monoclonal

Antigen

Other

Description

The antibody does stain especially simple epithelia. Only frozen sections can be used in immunoperoxidase and immunofluorescence tests. In immunoblotting experiments the antibody was shown to recognize only keratin no.18 (45 kDa).

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

M9

Vial size

1 mL

Concentration

15 µg/mL

Storage

4° C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Cytokeratin 18 Monoclonal Antibody (M9)(Product # MA1-35546) and a 48kDa band corresponding to Cytokeratin 18 was observed across the cell lines tested except IMR-32 and A-375. Whole cell extracts (30 µg lysate) of A549 (Lane 1), HeLa (Lane 2), Caco-2 (Lane 3), IMR-32 (Lane 4) and A-375 (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) 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 18 was performed using 70% confluent log phase Caco-2 cells. The cells were fixed with ice-cold acetone at 4°C for 5 minutes and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Cytokeratin 18 Monoclonal Antibody (M9) (Product # MA1-35546) at 1:100 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), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cytoskeletal localization. Panel e represents A-375 cells lacking cytoskeletal staining. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.