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Product Info Summary
| SKU: | A02277-3 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-EEF2K Antibody Picoband®
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. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
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.
Application & Images
Applications
A02277-3 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee
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
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human PC-3 whole cell, human U-87MG whole cell, human HepG2 whole cell, rat C6 whole cell
ICC/IF: HepG2 cell
FCM: SiHa cell
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.
Protein Target Info & Infographic
Gene/Protein Information For EEF2K (Source: Uniprot.org, NCBI)
Gene Name
EEF2K
Full Name
Eukaryotic elongation factor 2 kinase
Weight
75694 MW
Superfamily
protein kinase superfamily
Alternative Names
Arachidonate 12-lipoxygenase, 12S-type;12S-LOX;12S-lipoxygenase;1.13.11.31;Lipoxin synthase 12-LO;3.3.2.-;Platelet-type lipoxygenase 12;ALOX12;12LO, LOG12; 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
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on EEF2K, check out the EEF2K Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for EEF2K: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-EEF2K Antibody Picoband® (A02277-3)
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