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Phospho-EEF2K-S366 Rabbit pAb (AP0845)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse, Rat

Western blot analysis of lysates from HeLa cells, using Phospho-EEF2K-S366 Rabbit pAb (AP0845) at 1:1000 dilution. Hela cell lysate were treated by CIP (20ul CIP for each 400ul cell lysate) at 37℃ for 1 hour or treated by PMA/TPA (200nM) for 15 minutes after serum-starvation overnight.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% BSA.
Detection: ECL Basic Kit (RM00020).
Exposure time: 15s.

Western blot analysis of various lysates using Phospho-EEF2K-S366 Rabbit pAb (AP0845) at 1:1000 dilution. NIH/3T3 and C6 cells were treated by CIP(20uL/400ul) at 37℃ for 1 hour or PMA/TPA (200 nM) at 37℃ for 30 minutes after serum-starvation overnight.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 30s.

Western blot analysis of lysates from HeLa cells, using Phospho-EEF2K-S366 Rabbit pAb (A5404). Hela cell lysate were treated by CIP (20ul CIP for each 400ul cell lysate) at 37℃ for 1 hour or treated by PMA/TPA (200nM) for 15 minutes after serum-starvation overnight.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% BSA.
Detection: ECL Basic Kit (RM00020).
Exposure time: 15s.

Overview

Product name	Phospho-EEF2K-S366 Rabbit pAb
Catalog No.	AP0845
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG

Background

This gene encodes a highly conserved protein kinase in the calmodulin-mediated signaling pathway that links activation of cell surface receptors to cell division. This kinase is involved in the regulation of protein synthesis. It phosphorylates eukaryotic elongation factor 2 (EEF2) and thus inhibits the EEF2 function. The activity of this kinase is increased in many cancers and may be a valid target for anti-cancer treatment.

Immunogen information

Immunogen	A synthetic phosphorylated peptide around S366 of human EEF2K (NP_037434.1).
Sequence	TLSGS
Gene ID
Swiss Prot
Synonyms	eEF-2K; CaMKIII; HSU93850; Phospho-EEF2K-S366
Calculated MW	82kDa
Observed MW	110kDa

Applications

Reactivity	Human, Mouse, Rat
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:500 - 1:2000
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.01% thimerosal, 50% glycerol, pH7.3.
Key application	Western blotting
Positive samples	HeLa, HeLa treated by PMA, NIH/3T3 treated by PMA, C6 treated by PMA
Cellular location	cytoplasm, cytosol, dendritic spine, glutamatergic synapse, postsynaptic density.
Customer validation	WB(Homo sapiens)