Explore
Validate
Learn
Western blot
ELISAAntibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [2]
- Immunohistochemistry [2]
Submit
Validation data
Reference
Comment
Report error
- Product number
- MA5-15809 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- MEK6 Monoclonal Antibody (3H12B9)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Description
- MA5-15809 targets MAP2K6 in indirect ELISA, IHC and WB applications and shows reactivity with Human and mouse samples. The MA5-15809 immunogen is purified recombinant fragment of human MAP2K6 expressed in E. Coli. MA5-15809 detects MAP2K6 which has a predicted molecular weight of approximately 38kDa.
- Reactivity
- Human, Mouse
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 3H12B9
- 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.
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of MAP2K6 using MAP2K6 monoclonal antibody (Product # MA5-15809) in HepG2 (1), MCF-7 (2) and NIH/3T3 (3) cell lysate.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-MEK6 Monoclonal Antibody (3H12B9) (Product # MA5-15809) 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
- 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 (3H12B9) (Product # MA5-15809) 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.
- 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 (3H12B9) (Product # MA5-15809) 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
- Immunohistochemical analysis of paraffin-embedded ovarian cancer (left) and kidney cancer (right) using MAP2K6 monoclonal antibody (Product # MA5-15809) followed with DAB staining.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded ovarian cancer (left) and kidney cancer (right) using MAP2K6 monoclonal antibody (Product # MA5-15809) followed with DAB staining.
