Anti-EEF2K Antibody Picoband™

Product Name

Anti-EEF2K Antibody Picoband™

View all Eef2k Antibodies

SKU/Catalog Number

A02277-3

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-EEF2K Antibody Picoband™ catalog # A02277-3. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Rat.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-EEF2K Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A02277-3)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human EEF2K recombinant protein (Position: K162-A719).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A02277-3 is reactive to Eef2k in Human, Rat

Applications

A02277-3 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

82 kDa

Calculated molecular weight

75694 MW

Background of Eef2k

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. 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.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconstitution

Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5 μg/ml, Human, Rat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of EEF2K using anti-EEF2K antibody (A02277-3).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human PC-3 whole cell lysates,
Lane 2: human U-87MG whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: rat C6 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-EEF2K antigen affinity purified polyclonal antibody (Catalog # A02277-3) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for EEF2K at approximately 82 kDa. The expected band size for EEF2K is at 82 kDa.

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Figure 2. IF analysis of EEF2K using anti-EEF2K antibody (A02277-3).
EEF2K was detected in an immunocytochemical section of HepG2 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-EEF2K Antibody (A02277-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 3. Flow Cytometry analysis of SiHa cells using anti-EEF2K antibody (A02277-3).
Overlay histogram showing SiHa cells stained with A02277-3 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-EEF2K Antibody (A02277-3, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Gene/Protein Information For Eef2k (Source: Uniprot.org, NCBI)

Alternative Names

Calcium/calmodulin-dependent eukaryotic elongation factor 2 kinase; calcium/calmodulin-dependent eukaryotic elongation factor-2 kinase; EC 2.7.11; EC 2.7.11.20; eEF-2 kinase; eEF-2Kcalmodulin-dependent protein kinase III; elongation factor-2 kinase; eukaroytic elongation factor 2 kinase; eukaryotic elongation factor 2 kinase; eukaryotic elongation factor-2 kinase; HSU93850; MGC45041 Eef2k|C86191, eEF-2K|eukaryotic elongation factor-2 kinase|eukaryotic elongation factor 2 kinase|calcium/calmodulin-dependent eukaryotic elongation factor 2 kinase|eEF-2 kinase|eukaroytic elongation factor 2 kinase

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