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Purified anti-eEF2 Antibody

Pricing & Availability
Clone
4B3-G7-H5 (See other available formats)
Regulatory Status
RUO
Other Names
Eukaryotic Translation Elongation Factor 2, Polypeptidyl-tRNA Translocase, Epididymis Secretory Sperm Binding Protein
Isotype
Mouse IgG2b, κ
Ave. Rating
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Product Citations
publications
4B3-G7-H5_PURE_eEF2_Antibody_122221
Total cell lysates (15 µg total protein) from the indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with a 1:8000 dilution of purified anti-eEF2 antibody (clone 4B3-G7-H5) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-β-actin (Cat. No. 664803) was used as a loading control at a 1:25000 dilution (lower). Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • 4B3-G7-H5_PURE_eEF2_Antibody_122221
    Total cell lysates (15 µg total protein) from the indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with a 1:8000 dilution of purified anti-eEF2 antibody (clone 4B3-G7-H5) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-β-actin (Cat. No. 664803) was used as a loading control at a 1:25000 dilution (lower). Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • 4B3-G7-H5_PURE_eEF2_Antibody_2_122221
    ICC staining of purified anti-eEF2 antibody (clone 4B3-G7-H5) on HeLa cells. HeLa cells were fixed and permeabilized with 100% methanol and blocked with 5% FBS for 1 hour. The cells were then stained with a 1:100 dilution of mouse IgG2b, κ isotype control (Cat. No. 402202) (panel A) or the primary antibody (panel B), followed by incubation with 2.5 µg/mL of Alexa Fluor® 594 goat anti-mouse IgG (Cat. No. 405326) for 1 hour at room temperature. Nuclei were counterstained with DAPI, and the images were captured with a 60X objective.
  • 4B3-G7-H5_PURE_eEF2_Antibody_3_122221
    IHC staining of purified anti-eEF2 antibody (clone 4B3-G7-H5) on formalin-fixed paraffin-embedded human skin tissue. The tissue was incubated with a 1:100 dilution of the primary antibody overnight at 4°C, followed by incubation with 2.5 μg/mL of Alexa Fluor® 647 goat anti-mouse IgG (yellow) (Cat. No. 405322) for one hour at room temperature. Nuclei were counterstained with DAPI (blue), and the slide was mounted with ProLong™ Gold Antifade Mountant. The image was captured with a 10x objective. Scale bar = 50 μm
Cat # Size Price Quantity Check Availability Save
948501 25 µL £81
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948502 100 µL £201
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Description

eEF2 is a eukaryotic elongation factor required for GTP-dependent ribosomal translocation eEF2 and is negatively regulated by eEF2 kinase. In response to elevated cytosolic calcium, calmodulin binds to and stimulates eEF2 kinase activity, stimulating phosphorylation and inactivation of eEF2 elongation function. Hippocampal tissue from Alzheimer’s patients display hyperphosphorylation of eEF2 at threonine 56 compared to age-matched controls. Emerging studies also show that sub-psychomimetic doses of ketamine, which is used as a rapid-acting anti-depressant, result in a block of NMDAR and deactivation of eEF2 kinase, leading to reduced phosphorylation of eEF2 and increased eEF2-dependent translation of brain-derived neurotrophic factor.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Partial recombinant human eEF2 protein
Formulation
Phosphate-buffered solution, 0.5% BSA, 0.09% NaN3, bottle antibody at 0.1 mg/mL.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.1 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
IHC-P, ICC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 1:8000 dilution. For immunocytochemistry, a dilution range of 1:100 is recommended. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a dilution range of 1:200 is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

 

Application Notes

When used for western blotting, 4B3-G7-H5 dilutions exceeding 1:8000 may produce non-specific binding.

Clone 4B3-G7-H5 was tested for ICC using the following fixation-permeabilization methods: 4% PFA + Triton X-100, 4% PFA + methanol, and methanol-only. Of these methods, methanol-only produced the most robust eEF2 staining.

BioLegend offers two clones, W19410A and 4B3-G7-H5, that detect eEF2.

WB: Both W19410A and 4B3-G7-H5 detected eEF2 with a strong signal, but W19410A was more selective for eEF2. Both clones produced a stronger signal than CST’s antibody when the latter was used at the manufacturer’s recommended dilution.

ICC: W19410A and 4B3-G7-H5 performed similarly for the application.

IP: W19410A passed and 4B3-G7-H5 failed application testing.

IHC-P: W19410A and 4B3-G7-H5 both passed for the application, with 4B3-G7-H5 showing better selectivity.

 

RRID
AB_2910512 (BioLegend Cat. No. 948501)
AB_2910512 (BioLegend Cat. No. 948502)

Antigen Details

Structure
eEF2 is an 858 amino acid protein with a predicted molecular weight of 95 kD.
Distribution

Ubiquitously expressed/Cytosol

Function
Protein translation elongation factor
Antigen References
  1. Ryazanov AG, et al. 1991. FEBS Lett. 285:170-5.
  2. Autry AE, et al. 2011. Nature. 475:91-5.
  3. Beckelman BC, et al. 2019. J Clin Invest. 129:820-833.

 

Gene ID
1938 View all products for this Gene ID
UniProt
View information about eEF2 on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 1    Revision Date: 12/22/2021

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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