MA5-14088

antibody from Invitrogen Antibodies

Targeting: KRT18

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-14088

Provider

Invitrogen Antibodies

Product name

Cytokeratin 8/18 Monoclonal Antibody (5D3)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

MA5-14088 targets Cytokeratin 8/18 in immunohistochemistry (paraffin) applications and shows reactivity with Human samples. The MA5-14088 immunogen is cytokeratins purified from breast cancer MCF-7 cells.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

5D3

Vial size

500 µL

Concentration

Conc. Not Determined

Storage

4° C

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of Cytokeratin 8, 18 was performed using 70% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Cytokeratin 8, 18 (5D3) Mouse Monoclonal Antibody (Product # MA5-14088) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) a dilution of 1:2000 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). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Formalin-fixed, paraffin-embedded human skin stained with Keratin 8/18 antibody using peroxidase-conjugate and AEC chromogen. Note cytoplasmic staining of epithelial cells in the sweat glands.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of Cytokeratin 8, 18 was done on MCF7 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with Cytokeratin 8, 18 Mouse Monoclonal Antibody (MA5-14088, red histogram) or with mouse isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Rabbit Anti-Mouse Secondary Antibody (A11059) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10, 000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.