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Product Info Summary
| SKU: | PB9803 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-PKR/EIF2AK2 Antibody Picoband®
SKU/Catalog Number
PB9803
PB0855 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-PKR/EIF2AK2 Antibody Picoband® catalog # PB9803. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human. 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
Store 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 freeze-thaw cycles.
Cite This Product
Anti-PKR/EIF2AK2 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9803)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human PKR, different from the related mouse sequence by fifteen amino acids, and from the related rat sequence by thirteen amino acids.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
PB9803 is reactive to EIF2AK2 in Human
Observed Molecular Weight
69 kDa
Calculated molecular weight
62.1 kDa
Background of EIF2AK2
EIF2AK2 (Eukaryotic Translation Initiation Factor 2-Alpha Kinase 2), also called PKR, is an enzyme that in humans is encoded by the EIF2AK2 gene. Activation of EIF2AK2 allows the kinase to phosphorylate its natural substrate, the alpha subunit of eukaryotic protein synthesis initiation factor-2, leading to the inhibition of protein synthesis. By FISH analysis, Squire et al. (1993) assigned the EIF2AK2 gene to the boundary between chromosome 2p22-p21. Ben-Asouli et al. (2002) showed that human gamma-interferon mRNA uses local activation of PKR in the cell to control its own translation yield. IFNG mRNA was found to activate PKR through a pseudoknot in its 5-prime untranslated region. Taylor et al. (1999) studied the mechanism underlying the resistance of hepatitis C virus (HCV) to interferon. They demonstrated that the HCV envelope protein E2 contains a sequence identical with phosphorylation sites of the interferon-inducible protein kinase PKR and the translation initiation factor EIF2-alpha, a target of PKR. E2 inhibited the kinase activity of PKR and blocked its inhibitory effect on protein synthesis and cell growth.
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
PB9803 is guaranteed for 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, 0.1-0.5μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human Hela whole cell, human 293T whole cell, human MCF-7 whole cell, human K562 whole cell, human PANC-1 whole cell, human A431 whole cell, human U251 whole cell
ICC/IF: A549 cell
FCM: HEL cell
Validation Images & Assay Conditions
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Western blot analysis of PKR/EIF2AK2 using anti-PKR/EIF2AK2 antibody (PB9803).
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 Hela whole cell lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: human MCF-7 whole cell lysates,
Lane 4: human K562 whole cell lysates,
Lane 5: human PANC-1 whole cell lysates,
Lane 6: human A431 whole cell lysates,
Lane 7: human U251 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-PKR/EIF2AK2 antigen affinity purified polyclonal antibody (Catalog # PB9803) 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 PKR/EIF2AK2 at approximately 69 kDa. The expected band size for PKR/EIF2AK2 is at 62 kDa.
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Flow Cytometry analysis of HEL cells using anti-PKR/EIF2AK2 antibody (PB9803).
Overlay histogram showing HEL cells stained with PB9803 (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-PKR/EIF2AK2 Antibody (PB9803, 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.
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IF analysis of PKR/EIF2AK2 using anti-PKR/EIF2AK2 antibody (PB9803).
PKR/EIF2AK2 was detected in an immunocytochemical section of A549 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 2 μg/mL rabbit anti-PKR/EIF2AK2 Antibody (PB9803) overnight at 4°C. DyLight488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 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.
Specific Publications For Anti-PKR/EIF2AK2 Antibody Picoband® (PB9803)
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1 Customer Q&As for Anti-PKR/EIF2AK2 Antibody Picoband®
Question
We are currently using anti-PKR/EIF2AK2 antibody PB9803 for human tissue, and we are satisfied with the WB results. The species of reactivity given in the datasheet says human. Is it possible that the antibody can work on dog tissues as well?
T. Baker
Verified customer
Asked: 2015-12-25
Answer
The anti-PKR/EIF2AK2 antibody (PB9803) has not been validated for cross reactivity specifically with dog tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in dog you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2015-12-25
