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Product Info Summary
| SKU: | A00371-4 |
|---|---|
| Size: | 100 µg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-ATF4 Antibody Picoband®
SKU/Catalog Number
A00371-4
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-ATF4 Antibody Picoband® catalog # A00371-4. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, 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-ATF4 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00371-4)
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 ATF4 recombinant protein (Position: E10-E210).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00371-4 is reactive to ATF4 in Human, Mouse, Rat
Observed Molecular Weight
45-50 kDa
Calculated molecular weight
38.6 kDa
Background of ATF4
ATF4, Activating Transcription Factor 4, is also known as CREB2. ATF4 belongs to the large ATF/CREB family of transcription factors which bind DNA via their basic region and dimerize via their leucine zipper domain to form a variety of homo- and heterodimers to regulate gene transcription. It is identified that members of this family share significant sequence similarity within a leucine zipper DNA-binding motif and an adjacent basic region. The ATF4 gene is mapped to chromosome 22. Unlike CREB, which activates transcription from CRE-containing promoters, CREB2 functions as a specific repressor of CRE-dependent transcription. The transcriptional repressor activity resides within the C-terminal leucine zipper and basic domain region of the CREB2 protein.
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
A00371-4 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, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human HepG2 whole cell, human 293T whole cell, rat brain tissue, mouse brain tissue
ICC/IF: Hela cell
FCM: SiHa cell
Validation Images & Assay Conditions
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Western blot analysis of ATF4 using anti-ATF4 antibody (A00371-4).
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 HepG2 whole cell lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: rat brain tissue lysates,
Lane 4: mouse brain tissue 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-ATF4 antigen affinity purified polyclonal antibody (Catalog # A00371-4) 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 ATF4 at approximately 45-50 kDa. The expected band size for ATF4 is at 38,39,45-50 kDa.
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IF analysis of ATF4 using anti-ATF4 antibody (A00371-4) and anti-Tubulin Alpha antibody (M03989-3).
ATF4 was detected in immunocytochemical section of Hela cell. 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-ATF4 Antibody (A00371-4) and mouse anti-Tubulin Alpha antibody (M03989-3) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) and DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Flow Cytometry analysis of SiHa cells using anti-ATF4 antibody (A00371-4).
Overlay histogram showing SiHa cells stained with A00371-4 (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-ATF4 Antibody (A00371-4, 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 (Red line) was also used as a control.
Specific Publications For Anti-ATF4 Antibody Picoband® (A00371-4)
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