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Product Info Summary
| SKU: | A01172-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Monkey, Mouse |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-EIF2AK4 Antibody Picoband®
SKU/Catalog Number
A01172-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-EIF2AK4 Antibody Picoband® catalog # A01172-2. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Human, Monkey, Mouse. 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-EIF2AK4 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01172-2)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human EIF2AK4 recombinant protein (Position: Q23-K440).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A01172-2 is reactive to EIF2AK4 in Human, Monkey, Mouse
Observed Molecular Weight
180-200 kDa
Calculated molecular weight
186.9 kDa
Background of EIF2AK4
EIF2AK4 (Eukaryotic Translation Initiation Factor 2-Alpha Kinase 4), also called GCN2, is an enzyme that in humans is encoded by the EIF2AK4 gene. EIF2AK4 belongs to a family of kinases that phosphorylate the alpha subunit of eukaryotic translation initiation factor-2 to downregulate protein synthesis in response to varied cellular stresses. Hartz (2005) mapped the EIF2AK4 gene to chromosome 15q15.1 based on an alignment of the EIF2AK4 sequence with the genomic sequence. Berlanga et al. (1999) demonstrated that Gcn2 immunopurified from mouse liver extracts could phosphorylate rabbit Eif2 in vitro. Serum starvation increased the level of phosphorylated EIF2-alpha more than 2-fold in human embryonic kidney cells transfected with mouse Eif2ak4. Costa-Mattioli et al. (2005) reported a unique feature of hippocampal slices from Gcn2-null mice: in CA1, a single 100-Hz train induced a strong and sustained long-term potentiation (late LTP or L-LTP), which was dependent on transcription and translation.
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
A01172-2 is guaranteed for ELISA, Flow Cytometry, IHC, 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, 0.1-0.25μg/ml, Human, Mouse, Monkey
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human U87 whole cell, human HepG2 whole cell, monkey COS-7 whole cell, mouse brain tissue, mouse lung tissue, mouse ANA-1 whole cell
IHC: human mammary cancer tissue
FCM: THP-1 cell
Validation Images & Assay Conditions
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Western blot analysis of EIF2AK4 using anti-EIF2AK4 antibody (A01172-2).
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 50ug of sample under reducing conditions.
Lane 1: human U87 whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: monkey COS-7 whole cell lysates,
Lane 4: mouse brain tissue lysates,
Lane 5: mouse lung tissue lysates,
Lane 6: mouse ANA-1 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-EIF2AK4 antigen affinity purified polyclonal antibody (Catalog # A01172-2) at 0.25 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 1:10000 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 EIF2AK4 at approximately 180-200KD. The expected band size for EIF2AK4 is at 187KD.
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IHC analysis of EIF2AK4 using anti-EIF2AK4 antibody (A01172-2).
EIF2AK4 was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-EIF2AK4 Antibody (A01172-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
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Flow Cytometry analysis of THP-1 cells using anti-EIF2AK4 antibody (A01172-2).
Overlay histogram showing THP-1 cells stained with A01172-2 (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-EIF2AK4 Antibody (A01172-2,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-EIF2AK4 Antibody Picoband® (A01172-2)
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