Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
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Validation data
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- Product number
- PA5-17416 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- MEK6 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- It is not recommended to aliquot this antibody.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of NhpK6 in extracts from HeLa and NIH/3T3 cells using NhpK6 polyclonal antibody (Product # PA5-17416).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-MEK6 Polyclonal Antibody (Product # PA5-17416) 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-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
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 Polyclonal Antibody (Product # PA5-17416) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 (Product # A27034, 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.