14-9815-82

antibody from Invitrogen Antibodies

Targeting: KRT18

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

14-9815-82

Provider

Invitrogen Antibodies

Product name

Cytokeratin 18 Monoclonal Antibody (LDK18), eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The monoclonal antibody LDK18 recognizes human Cytokeratin 18. Cytokeratin 18 is member of the large family of intermediate filaments found within epithelial cells. The family of cytokeratins is made up of type I (acidic) and type II (basic) members which form heterodimers by pairing an acidic with a basic member to compose the keratin filament. Cytokeratin 18 (type I) is most commonly found associated with cytokeratin 8 (type II). Because many tumors of epithelial origin retain expression of keratin proteins, cytokeratin 18 has been used as a tumor marker to identify both tumors and metastases. In addition, cytokeratin 18 has been shown to be expressed in highly proliferative retinal pigment epithelium cells.

Antibody clone number

LDK18

Concentration

0.5 mg/mL

References

Submitted references

The human keratins: biology and pathology.

Moll R, Divo M, Langbein L
Histochemistry and cell biology 2008 Jun;129(6):705-33

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The cytoskeletal elements of human retinal pigment epithelium: in vitro and in vivo.

McKechnie NM, Boulton M, Robey HL, Savage FJ, Grierson I
Journal of cell science 1988 Oct;91 ( Pt 2):303-12

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Monoclonal cytokeratin antibodies that distinguish simple from stratified squamous epithelia: characterization on human tissues.

Debus E, Weber K, Osborn M
The EMBO journal 1982;1(12):1641-7

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Cytokeratin 18 Monoclonal Antibody (LDK18), eBioscience™(Product # 14-9815-82) 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 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

Immunocytochemistry of fixed MCF-7 cells stained with 1 µg/mL Mouse IgG1 K Isotype Control Purified (left) or 1 µg/mL Anti-Human Cytokeratin 18 Purified (right) followed by F (ab')2 Anti-Mouse IgG eFluor® 570 (Product # 41-4010-82).Nuclei are stained with DAPI.

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 (LDK18), eBioscience™ (Product # 14-9815-82) at 5 µg/mL 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. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.