MA5-15808

antibody from Invitrogen Antibodies

Targeting: MAP2K6 MAPKK6, MEK6, MKK6, PRKMK6, SAPKK3

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-15808

Provider

Invitrogen Antibodies

Product name

MEK6 Monoclonal Antibody (3H12C8)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Reactivity

Human, Mouse

Host

Mouse

Isotype

IgG

Antibody clone number

3H12C8

Vial size

100 µL

Concentration

Conc. Not Determined

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-MEK6 Monoclonal Antibody (3H12C8) (Product # MA5-15808) and a 37 kDa band corresponding to MEK6 was observed across all the cell lines and tissues tested except HCT 116, SK-OV-3 and K-562. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), HEK-293 (Lane 2), HeLa (Lane 3), HCT 116 (Lane 4), NIH/3T3 (Lane 5), SK-OV-3 (Lane 6), K-562 (Lane 7), tissue extracts of Mouse cerebellum (Lane 8), Mouse Skeletal Muscle (Lane 9), Mouse White Adipose (Lane 10) and Mouse Colon (Lane 11) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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). A band at ~ 25 kDa was observed in all the tissues which corresponds to circulating tissue IgG.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of MAP2K6 using MAP2K6 monoclonal antibody (Product # MA5-15808) in HepG2 (1), MCF-7 (2) and NIH/3T3 (3) cell lysate.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of MEK6 was performed using 70% confluent log phase HeLa and HCT 116 cells. The cells were fixed with 4% Paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with MEK6 Monoclonal Antibody (3H12C8) (Product # MA5-15808) 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™ 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). Panel d represents the merged image showing nuclear and cytoplasmic localization. Panel e represents HCT 116 cells having no expression of MEK6. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of HeLa cells using MAP2K6 monoclonal antibody (Product # MA5-15808) (Green). Blue: DRAQ5 fluorescent DNA dye.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometric analysis of HeLa cells using MAP2K6 monoclonal antibody (Product # MA5-15808) (green) and negative control (purple).