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SKU:PB9131
Pack Size:100μg/vial
Sample Size:30ug for $99, contact us for details
Clonality:Polyclonal
Application:Flow Cytometry, IHC, ICC, WB
Price: $240.00
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Overview

Product Name Anti-ATF2 Picoband™ Antibody
SKU/Catalog Number PB9131
Description Rabbit IgG polyclonal antibody for Cyclic AMP-dependent transcription factor ATF-2(ATF2) detection. Tested with WB, IHC-P, IHC-F, ICC, FCM in Human;Mouse;Rat.
Cite This Product Anti-ATF2 Picoband™ Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9131)
Replacement Item This antibody may replace the following items: sc-101635|sc-101636|sc-101637|sc-101638|sc-13147|sc-135686|sc-187|sc-187-X|sc-242|sc-242-X|sc-32551|sc-328|sc-328-G|sc-398555|sc-52941|sc-6233|sc-6233-X|sc-7982|sc-7982-R|sc-7982-X|sc-8398|sc-8398-X|sc-9010 from Santa Cruz Biotechnology.
Host Rabbit
Isotype N/A
Validated Species Human, Mouse, Rat
Application Flow Cytometry, IHC, ICC, WB

*Our Boster Guarantee covers the use of this product in the above tested applications.

**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.

Recommended Detection Systems Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P), IHC(F) and ICC.
*Blocking peptide can be purchased at $50. Contact us for more information
**Boster also offers various secondary antibodies for Immunoflourescecne and IHC. Take advantage of the buy 1 primary antibody get 1 secondary antibody for free promotion for the entire year 2017!
Immunogen E.coli-derived human ATF2 recombinant protein (Position: E93-E450). Human ATF2 shares 99% amino acid (aa) sequence identity with both mouse and rat ATF2.
Cross Reactivity No cross reactivity with other proteins
Pack Size 100μg/vial

Properties

Clonality Polyclonal
Form Lyophilized
Contents Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
*carrier free antibody available upon request.
Concentration Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Storage At -20˚C for one year. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for a longer time.Avoid repeated freezing and thawing.
Purification Immunogen affinity purified.
Isotype N/A

Protein Target Info (Source: Uniprot.org)

You can check the tissue specificity below for information on selecting positive and negative control.

Gene Name ATF2
Protein Name Cyclic AMP-dependent transcription factor ATF-2
Molecular Weight 54537 MW
Protein Function Transcriptional activator which regulates the transcription of various genes, including those involved in anti- apoptosis, cell growth, and DNA damage response. Dependent on its binding partner, binds to CRE (cAMP response element) consensus sequences (5'-TGACGTCA-3') or to AP-1 (activator protein 1) consensus sequences (5'-TGACTCA-3'). In the nucleus, contributes to global transcription and the DNA damage response, in addition to specific transcriptional activities that are related to cell development, proliferation and death. In the cytoplasm, interacts with and perturbs HK1- and VDAC1-containing complexes at the mitochondrial outer membrane, thereby impairing mitochondrial membrane potential, inducing mitochondrial leakage and promoting cell death. The phosphorylated form (mediated by ATM) plays a role in the DNA damage response and is involved in the ionizing radiation (IR)-induced S phase checkpoint control and in the recruitment of the MRN complex into the IR-induced foci (IRIF). Exhibits histone acetyltransferase (HAT) activity which specifically acetylates histones H2B and H4 in vitro. In concert with CUL3 and RBX1, promotes the degradation of KAT5 thereby attenuating its ability to acetylate and activate ATM. Can elicit oncogenic or tumor suppressor activities depending on the tissue or cell type. .
Tissue Specificity Ubiquitously expressed, with more abundant expression in the brain.
Sequence Similarities Belongs to the bZIP family. ATF subfamily.
Subcellular Localization Nucleus. Cytoplasm. Mitochondrion outer membrane. Shuttles between the cytoplasm and the nucleus and heterodimerization with JUN is essential for the nuclear localization. Localization to the cytoplasm is observed under conditions of cellular stress and in disease states. Localizes at the mitochondrial outer membrane in response to genotoxic stress. Phosphorylation at Thr-52 is required for its nuclear localization and negatively regulates its mitochondrial localization. Co- localizes with the MRN complex in the IR-induced foci (IRIF).
Uniprot ID P15336
Alternative Names Cyclic AMP-dependent transcription factor ATF-2;cAMP-dependent transcription factor ATF-2;2.3.1.48 ;Activating transcription factor 2;Cyclic AMP-responsive element-binding protein 2;CREB-2;cAMP-responsive element-binding protein 2;HB16;Histone acetyltransferase ATF2;cAMP response element-binding protein CRE-BP1;ATF2;CREB2, CREBP1;
Research Areas |epigenetics and nuclear signaling|transcription|domain families|hlh / leucine zipper| immunology|innate immunity|tlr signaling|
*if product is indicated to react with multiple species, protein info is based on the human gene.

Background for Cyclic AMP-dependent transcription factor ATF-2

ATF2, also known as Activating transcription factor 2, is a protein that in humans is encoded by the ATF2 gene. It is mapped to 2q31.1. This gene encodes a transcription factor that is a member of the leucine zipper family of DNA-binding proteins. This protein binds to the cAMP-responsive element (CRE), an octameric palindrome. The protein forms a homodimer or heterodimer with c-Jun and stimulates CRE-dependent transcription. The protein is also a histone acetyltransferase (HAT) that specifically acetylates histones H2B and H4 in vitro, thus, it may represent a class of sequence-specific factors that activate transcription by direct effects on chromatin components. Additional transcript variants have been identified but their biological validity has not been determined.

Anti-ATF2 Picoband™ Antibody Images

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Anti-ATF2 Picoband™ Antibody
Figure 1. Western blot analysis of ATF2 using anti-ATF2 antibody (PB9131).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours.
Lane 1: Recombinant Human ATF2 Protein 0.5ng.
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-ATF2 antigen affinity purified polyclonal antibody (Catalog # PB9131) 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 1:10000 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 ATF2 at approximately 49KD. The expected band size for ATF2 is at 49KD.
Anti-ATF2 Picoband™ Antibody
Figure 2. IHC analysis of ATF2 using anti-ATF2 antibody (PB9131).
ATF2 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-ATF2 Antibody (PB9131) 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.
Anti-ATF2 Picoband™ Antibody
Figure 3. IHC analysis of ATF2 using anti-ATF2 antibody (PB9131).
ATF2 was detected in paraffin-embedded section of mouse brain 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-ATF2 Antibody (PB9131) 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.
Anti-ATF2 Picoband™ Antibody
Figure 4. IHC analysis of ATF2 using anti-ATF2 antibody (PB9131).
ATF2 was detected in paraffin-embedded section of rat brain 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-ATF2 Antibody (PB9131) 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.
Anti-ATF2 Picoband™ Antibody
Figure 5. IHC analysis of ATF2 using anti-ATF2 antibody (PB9131).
ATF2 was detected in frozen section of rat brain 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-ATF2 Antibody (PB9131) 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.
Anti-ATF2 Picoband™ Antibody
Figure 6. IHC analysis of ATF2 using anti-ATF2 antibody (PB9131).
ATF2 was detected in frozen section of mouse brain 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-ATF2 Antibody (PB9131) 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.
Anti-ATF2 Picoband™ Antibody
Figure 7. Flow Cytometry analysis of K562 cells using anti-ATF2 antibody (PB9131).
Overlay histogram showing K562 cells stained with PB9131 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATF2 Antibody (PB9131,1μg/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 (Red line) was also used as a control.
Anti-ATF2 Picoband™ Antibody
Figure 8. Flow Cytometry analysis of HepG2 cells using anti-ATF2 antibody (PB9131).
Overlay histogram showing HepG2 cells stained with PB9131 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATF2 Antibody (PB9131,1μg/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 (Red line) was also used as a control.
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FAQs

Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugation
A: Yes, please contact us at support@bosterbio.com for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugation
A: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact support@bosterbio.com
Q: What should I use for negative control?
A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?
A: You can find the immunogen sequence under "Immunogen" and clonality in the product name.
Q: What is the expected band size? Why is it different than the observed band size?
A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:
1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands

3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.

4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,
Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?
A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?
A: Check our protocols under the tech support tab.
Q: What are some alternative names that could be used to describe this product?
A: Some common names include but are not limited to atf 2 antibody, atf2 antibody, atf-2 antibody