EEF2K Recombinant Rabbit Monoclonal Antibody [PSH11-67]
Catalog# HA723363
EEF2K Recombinant Rabbit Monoclonal Antibody [PSH11-67]
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WB
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IF-Cell
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
EEF2K Recombinant Rabbit Monoclonal Antibody [PSH11-67]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within human EEF2K aa 476-725.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, FC
Target Molecular Weight
Predicted band size: 82 kDa
Positive Control
HeLa cell lysate, 293T cell lysate, K-562 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, Mouse ovary tissue lysate, Rat ovary tissue lysate, HeLa-parental cell lysate, HeLa.
Conjugation
unconjugated
Clone Number
PSH11-67
Reactivity Data
| WB | IF-Cell | FC | |
|---|---|---|---|
| Human |
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|
|
| Mouse |
|
|
|
| Rat |
|
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:5,000-1:10,000
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IF-Cell
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1:100
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FC
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1:1,000
Target
Function
Eukaryotic elongation factor-2 kinase (eEF-2 kinase or eEF-2K), also known as calmodulin-dependent protein kinase III (CAMKIII) and calcium/calmodulin-dependent eukaryotic elongation factor 2 kinase, is an enzyme that in humans is encoded by the EEF2K gene. eEF-2 kinase is a highly conserved protein kinase in the calmodulin-mediated signaling pathway that links multiple up-stream signals to the regulation of protein synthesis. It phosphorylates eukaryotic elongation factor 2 (EEF2) and thus inhibits the EEF2 function.
Background References
1. Klupt KA et al. eEF2K Inhibitor Design: The Progression of Exemplary Structure-Based Drug Design. Molecules. 2023 Jan
2. Chen X et al. eEF2K promotes PD-L1 stabilization through inactivating GSK3beta in melanoma. J Immunother Cancer. 2022 Mar
Subcellular Location
cytoplasm, cytosol, dendritic spine, glutamatergic synapse, postsynaptic density.
Synonyms
C86191 antibody
Calcium/calmodulin dependent eukaryotic elongation factor 2 antibody
Calcium/calmodulin dependent eukaryotic elongation factor 2 kinase antibody
Calcium/calmodulin-dependent eukaryotic elongation factor 2 kinase antibody
Calmodulin dependent protein kinase III antibody
cb365 antibody
EC 2.7.11.20 antibody
eEF 2 kinase antibody
eEF 2K antibody
eEF-2 kinase antibody
ExpandImages
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Western blot analysis of EEF2K on different lysates with Rabbit anti-EEF2K antibody (HA723363) at 1/10,000 dilution.
Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: 293T cell lysate (20 µg/Lane)
Lane 3: K-562 cell lysate (20 µg/Lane)
Lane 4: NIH/3T3 cell lysate (20 µg/Lane)
Lane 5: C6 cell lysate (20 µg/Lane)
Lane 6: Mouse ovary tissue lysate (40 µg/Lane)
Lane 7: Rat ovary tissue lysate (40 µg/Lane)
Predicted band size: 82 kDa
Observed band size: 82-90 kDa
Exposure time: 25 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723363) at 1/10,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Knockdown (KD)
Western blot analysis of EEF2K on different lysates with Rabbit anti-EEF2K antibody (HA723363) at 1/5,000 dilution.
Lane 1: HeLa-parental cell lysate
Lane 2: HeLa-EEF2K KD cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 82 kDa
Observed band size: 82-90 kDa
Exposure time: 10 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723363) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of HeLa cells labeling EEF2K with Rabbit anti-EEF2K antibody (HA723363) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-EEF2K antibody (HA723363) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Flow cytometric analysis of HeLa cells labeling EEF2K.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA723363, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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