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Purified anti-eEF2 Antibody

Pricing & Availability
Clone
W19410A (See other available formats)
Regulatory Status
RUO
Other Names
Eukaryotic Translation Elongation Factor 2, Polypeptidyl-tRNA Translocase, Epididymis Secretory Sperm Binding Protein
Isotype
Rat IgG2b, κ
Ave. Rating
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Product Citations
publications
1_W19410A_PURE_eEF2_Antibody_WB_092921.png
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 W19410A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-β-actin antibody (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.
  • 1_W19410A_PURE_eEF2_Antibody_WB_092921.png
    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 W19410A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-β-actin antibody (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.
  • 2_W19410A_PURE_eEF2_Antibody_ICC_092921.png
    ICC staining of purified anti-eEF2 antibody (clone W19410A) on eEF2-depleted HeLa cells. HeLa cells treated with control siRNA (panel A) or siRNA targeting eEF2 (panel B) 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 the primary antibody, followed by incubation with 2.5 µg/mL of Alexa Fluor® 594 anti-goat anti-rat IgG antibody (Cat. No. 405422) for 1 hour at room temperature. Nuclei were counterstained with DAPI, and the images were captured with a 60X objective.
  • 3_W19410A_PURE_eEF2_Antibody_IHC_092921.png
    IHC staining of purified anti-eEF2 antibody (clone W19410A) 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.
  • 4_W19410A_PURE_eEF2_Antibody_IP_092921.png
    Whole cell extracts (250 µg total protein) prepared from HeLa cells were immunoprecipitated overnight with 2.5 µg of purified rat IgG2b, κ Isotype ctrl antibody (Cat. No. 400602) or 25 µL of purified anti-eEF2 antibody (clone W19410A). The resulting IP fractions and whole cell extract input (6%) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with a control antibody against a separate epitope of eEF2. Lane M: Molecular weight marker.
Cat # Size Price Quantity Check Availability Save
947601 25 µL 95€
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947602 100 µL 235€
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Description

eEF2 is a eukaryotic elongation factor required for GTP-dependent ribosomal translocation 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
Rat
Immunogen
Partial recombinant human eEF2 protein
Formulation
Phosphate buffer, 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
ICC, IHC-P, IP, KO/KD-WB - 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 a 1:8000 dilution. For immunocytochemistry, a dilution of 1:100 dilution is recommended. For immunohistochemistry, a dilution of 1:100 dilution is suggested. For immunoprecipitation, the suggested use of this reagent is 25 µg/test. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

When used for western blotting, W19410A antibody concentrations exceeding 0.0125 µg/mL may produce non-specific binding.

Clone W9410A was tested for ICC using the following fixation-permeabilization methods: 4% PFA + Triton X-100, 4% PFA + methanol, and methanol-only. All of these methods were compatible with eEF2 staining.

BioLegend offers two sizes (25 and 100 µL0 for W19410A. If using this clone for immunoprecipitation, we recommend purchasing the 100 µL size to ensure sufficient material for the application.

RRID
AB_2904459 (BioLegend Cat. No. 947601)
AB_2904459 (BioLegend Cat. No. 947602)

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
Biology Area
Cell Biology, Protein Synthesis
Antigen References

1. Ryazanov AG, et al. 1991. FEBS. Lett. 285: 170.
2. Autry AE, et al. 2011. Nature. 475: 91.
3. Beckelman BC, et al. 2019. J. Clin. Invest. 129: 820.

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

Other Formats

View All eEF2 Reagents Request Custom Conjugation
Description Clone Applications
Purified anti-eEF2 Antibody W19410A WB,IHC-P,ICC,IP,KO/KD-WB
Go To Top Version: 1    Revision Date: 09.29.2021

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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