Phospho-EEF2 (T56) Recombinant Rabbit Monoclonal Antibody [PSH06-36]
Catalog# HA722670
Phospho-EEF2 (T56) Recombinant Rabbit Monoclonal Antibody [PSH06-36]
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WB
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IHC-P
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IF-Cell
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
Phospho-EEF2 (T56) Recombinant Rabbit Monoclonal Antibody [PSH06-36]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic phosphopeptide corresponding to residues surrounding Thr56 of human EEF2.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, IF-Cell
Target Molecular Weight
Predicted band size: 95 kDa
Positive Control
HeLa starved for 24 hours then add 10μM forskolin for 1 hour cell lysate, NIH/3T3 cell lysate, NIH/3T3 starved for 24 hours then add 10μM forskolin for 1 hour cell lysate, PC-12 treated with 20μM forskolin for 1 hour then add 100nM Calyculin A for 30 minutes cell lysate, HepG2 cell lysate, rat liver tissue, HeLa cells starved for 24 hours then add 10μM forskolin for 1 hour, PC-12 cells treated with 20μM forskolin for 1 hour then add 100nM Calyculin A for 30 minutes.
Conjugation
unconjugated
Clone Number
PSH06-36
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
PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:2,000
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IHC-P
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1:500
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IF-Cell
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1:100
Target
Function
Eukaryotic elongation factor 2 is a protein that in humans is encoded by the EEF2 gene. It is the archaeal and eukaryotic counterpart of bacterial EF-G. This gene encodes a member of the GTP-binding translation elongation factor family. This protein is an essential factor for protein synthesis. It promotes the GTP-dependent translocation of the ribosome. This protein is completely inactivated by EF-2 kinase phosphorylation. aEF2/eEF2 found in most archaea and eukaryotes, including humans, contains a post translationally modified histidine diphthamide. It is the target of diphtheria toxin (from Corynebacterium diphtheriae), and exotoxin A (from Pseudomonas aeruginosa). The inactivation of EF-2 by toxins inhibits protein production in the host, causing symptoms due to loss of function in affected cells.
Background References
1. Chen Q et al. Inhibition of LDHA to induce eEF2 release enhances thrombocytopoiesis. Blood. 2022 May
2. Li W et al. Fluoxetine regulates eEF2 activity (phosphorylation) via HDAC1 inhibitory mechanism in an LPS-induced mouse model of depression. J Neuroinflammation. 2021 Feb
Subcellular Location
Cytoplasm, Nucleus.
Synonyms
EEF 2 antibody
Eef2 antibody
EF-2 antibody
EF2 antibody
EF2_HUMAN antibody
Elongation factor 2 antibody
Eukaryotic translation elongation factor 2 antibody
Polypeptidyl tRNA translocase antibody
SCA26 antibody
Images
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☑ Cell treatment (CT)
Western blot analysis of Phospho-EEF2 (T56) on different lysates with Rabbit anti-Phospho-EEF2 (T56) antibody (HA722670) at 1/2,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: HeLa starved for 24 hours then add 10μM forskolin for 1 hour cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: NIH/3T3 starved for 24 hours then add 10μM forskolin for 1 hour cell lysate
Lane 5: PC-12 cell lysate
Lane 6: PC-12 treated with 20μM forskolin for 1 hour then add 100nM Calyculin A for 30 minutes cell lysate
Lane 7: HepG2 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 95 kDa
Observed band size: 95 kDa
Exposure time: 3 minutes; ECL: K1802;
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 (HA722670) at 1/2,000 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. -
Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-Phospho-EEF2 (T56) antibody (HA722670) at 1/500 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722670) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunocytochemistry analysis of HeLa cells starved for 24 hours then add 10μM forskolin for 1 hour labeling Phospho-EEF2 (T56) with Rabbit anti-Phospho-EEF2 (T56) antibody (HA722670) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Phospho-EEF2 (T56) antibody (HA722670) 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 (M1305-2, 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. -
☑ Cell treatment (CT)
Immunocytochemistry analysis of PC-12 cells treated with 20μM forskolin for 1 hour then add 100nM Calyculin A for 30 minutes labeling Phospho-EEF2 (T56) with Rabbit anti-Phospho-EEF2 (T56) antibody (HA722670) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Phospho-EEF2 (T56) antibody (HA722670) 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 (M1305-2, 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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"