GTX24760

antibody from GeneTex

Targeting: MAP2K4 JNKK1, MEK4, MKK4, PRKMK4, SERK1

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

GTX24760

Provider

GeneTex

Proper citation

GeneTex Cat#GTX24760, RRID:AB_380410

Product name

SEK1 / MKK4 (phospho Ser257/Thr261) antibody

Antibody type

Polyclonal

Reactivity

Human, Mouse, Xenopus

Host

Rabbit

Storage

Aliquot and store at -20°C or -80¢XC. Avoid repeated freeze / thaw cycles.

Western blot

Supportive validation

Submitted by

GeneTex (provider)

Main image

Experimental details

GST-tagged fusion protein expressing MEK4 was spiked into background extracts and resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer overnight at 4°C, then incubated with the MEK4 [pSpT257/261] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1), the non-phosphopeptide corresponding to the phoshpopeptide immunogen (2), a generic phosphoserine-containing peptide (3), a generic phosphothreonine-containing peptide (4), or the phosphopeptide immunogen (5). After washing, the membrane was incubated with goat F(ab)2 anti-rabbit IgG alkaline phosphatase and signals were detected. Phosphatase Stripping: Extracts were prepared as above. The membrane was left untreated (6, 8) or treated (7, 9) with lambda phosphatase, then incubated with either the MEK4 [pSpT257/261] antibody (6, 7) or a MEK4 pan antibody (8, 9) for two hours at room temperature, and developed using the same procedure as above. The data show that only the phosphopeptide corresponding to MEK4 [pSpT257/261] blocks the antibody signal, and that phosphatase stripping eliminates only the phospho-specific signal, demonstrating the specificity of the antibody. Please note that the GST-tagged fusion protein expressing MEK4 runs at ~67 kDa.