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Product Info Summary
| SKU: | PB9130 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-ATF1 Antibody Picoband®
SKU/Catalog Number
PB9130
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-ATF1 Antibody Picoband® catalog # PB9130. Tested in Flow Cytometry, ICC/IF, IHC, WB applications. This antibody reacts with HumanThe 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-ATF1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9130)
Host
Rabbit
Contents
Each vial contains antibody formulated with stabilizing components, 0.9 mg NaCl, 0.2 mg Na2HPO4, and 0.05 mg NaN3.
*This antibody is supplied in a stabilized formulation.
Compatibility with conjugation reactions depends on the chemistry of the conjugation method used.
For conjugation methods that are not compatible with the stabilizing components present in this formulation, a carrier-free antibody format is required.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human ATF1 recombinant protein (Position: M1-V271). Human ATF1 shares 91% amino acid (aa) sequence identity with mouse ATF1.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9130 is reactive to ATF1 in Human
Observed Molecular Weight
38 kDa
Calculated molecular weight
29.2 kDa
Background of ATF1
ATF1, also known as activating transcription factor 1, is a protein that in humans is encoded by the ATF1 gene. It is mapped to 12q13.12. This gene encodes an activating transcription factor, which belongs to the ATF subfamily and bZIP (basic-region leucine zipper) family. It influences cellular physiologic processes by regulating the expression of downstream target genes, which are related to growth, survival, and other cellular activities. This protein is phosphorylated at serine 63 in its kinase-inducible domain by serine/threonine kinases, cAMP-dependent protein kinase A, calmodulin-dependent protein kinase I/II, mitogen- and stress-activated protein kinase and cyclin-dependent kinase 3 (cdk-3). Its phosphorylation enhances its transactivation and transcriptional activities, and enhances cell transformation.
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
PB9130 is guaranteed for Flow Cytometry, IF, IHC, 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
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human Hela whole cell, human 293T whole cell, human K562 whole cell
IHC: human intestinal cancer tissue
ICC/IF: SiHa cell
FCM: SiHa cell
Validation Images & Assay Conditions
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Western blot analysis of ATF1 using anti-ATF1 antibody (PB9130).
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 K562 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-ATF1 antigen affinity purified polyclonal antibody (Catalog # PB9130) 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 ATF1 at approximately 38 kDa. The expected band size for ATF1 is at 38 kDa.
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IF analysis of ATF1 using anti-ATF1 antibody (PB9130).
ATF1 was detected in an immunocytochemical section of SiHa 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-ATF1 Antibody (PB9130) overnight at 4°C. Fluoro488 Conjugated Goat Anti-Rabbit IgG (BA1127) was 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-ATF1 antibody (PB9130).
Overlay histogram showing SiHa cells stained with PB9130 (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-ATF1 Antibody (PB9130,1μg/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.
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IHC analysis of ATF1 using anti-ATF1 antibody (PB9130).
ATF1 was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ATF1 Antibody (PB9130) 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.
Specific Publications For Anti-ATF1 Antibody Picoband® (PB9130)
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1 Customer Q&As for Anti-ATF1 Antibody Picoband®
Question
We are currently using anti-ATF1 antibody PB9130 for rat tissue, and we are content with the WB results. The species of reactivity given in the datasheet says human, rat. Is it likely that the antibody can work on zebrafish tissues as well?
Verified Customer
Verified customer
Asked: 2020-02-20
Answer
The anti-ATF1 antibody (PB9130) has not been validated for cross reactivity specifically with zebrafish tissues, though 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 zebrafish you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2020-02-20
