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Product Info Summary
| SKU: | A00225-3 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, WB |
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Product info
Product Name
Anti-Aryl hydrocarbon Receptor/Ahr Antibody Picoband®
SKU/Catalog Number
A00225-3
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Aryl hydrocarbon Receptor/Ahr Antibody Picoband® catalog # A00225-3. Tested in ELISA, Flow Cytometry, WB applications. This antibody reacts with 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
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-Aryl hydrocarbon Receptor/Ahr Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00225-3)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived rat Aryl hydrocarbon Receptor/Ahr recombinant protein (Position: R15-Q196).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00225-3 is reactive to Ahr in Mouse, Rat
Observed Molecular Weight
110 kDa
Calculated molecular weight
96.2 kDa
Background of Ahr
AHR (aryl hydrocarbon receptor), also called bHLHe76, is a member of the family of basic helix-loop-helix transcription factors. AhR is a cytosolic transcription factor that is normally inactive, bound to several co-chaperones. The AHR gene is mapped on 7p21.1. Estrogenic actions of AHR agonists were detected in wildtype ovariectomized mouse uteri, but were absent in Ahr -/- or Er-alpha -/- ovariectomized mice. Complex assembly and ubiquitin ligase activity of CUL4B (AHR) in vitro and in vivo are dependent on the AHR ligand. In the CUL4B (AHR) complex, ligand-activated AHR acts as a substrate-specific adaptor component that targets sex steroid receptors for degradation. Cd4-positive cells from mice lacking Ahr developed Th17 responses but failed to produce Il22 and did not show enhanced Th17 development. Activation of Ahr during induction of EAE accelerated disease onset and increased pathology in wildtype mice, but not in Ahr -/- mice. The TDO-AHR pathway is active in human brain tumors and is associated with malignant progression and poor survival. Ahr activity within ROR-gamma-t-positive ILC could be induced by dietary ligands such as those contained in vegetables of the family Brassicaceae.
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
A00225-3 is guaranteed for ELISA, Flow Cytometry, 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, 0.25-0.5μg/ml, Mouse, Rat
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Rat
ELISA, 0.1-0.5μg/ml,
Positive Control
WB: rat liver tissue, rat PC-12 whole cell, rat C6 whole cell, rat RH35 whole cell, mouse liver tissue
FCM: Neuro-2a cell
Validation Images & Assay Conditions
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Western blot analysis of Ahr using anti-Ahr antibody (A00225-3).
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: rat liver tissue lysates,
Lane 2: rat PC-12 whole cell lysates,
Lane 3: rat C6 whole cell lysates,
Lane 4: rat RH35 whole cell lysates,
Lane 5: mouse liver tissue 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-Ahr antigen affinity purified polyclonal antibody (Catalog # A00225-3) 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 Ahr at approximately 110KD. The expected band size for Ahr is at 96KD.
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Flow Cytometry analysis of Neuro-2a cells using anti-Ahr antibody (A00225-3).
Overlay histogram showing Neuro-2a cells stained with A00225-3 (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-Ahr Antibody (A00225-3, 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-Aryl hydrocarbon Receptor/Ahr Antibody Picoband® (A00225-3)
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4 Customer Q&As for Anti-Aryl hydrocarbon Receptor/Ahr Antibody Picoband®
Question
We are currently using anti-Aryl hydrocarbon Receptor/Ahr antibody A00225-3 for mouse tissue, and we are satisfied with the WB results. The species of reactivity given in the datasheet says mouse, rat. Is it likely that the antibody can work on feline tissues as well?
Verified Customer
Verified customer
Asked: 2020-02-04
Answer
The anti-Aryl hydrocarbon Receptor/Ahr antibody (A00225-3) has not been validated for cross reactivity specifically with feline 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 feline you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2020-02-04
Question
My colleagues were satisfied with the WB result of your anti-Aryl hydrocarbon Receptor/Ahr antibody. However we have seen positive staining in placenta cytoplasm. using this antibody. Is that expected? Could you tell me where is AHR supposed to be expressed?
Verified Customer
Verified customer
Asked: 2019-07-01
Answer
According to literature, placenta does express AHR. Generally AHR expresses in cytoplasm. Regarding which tissues have AHR expression, here are a few articles citing expression in various tissues:
Liver, Pubmed ID: 8393992
Placenta, Pubmed ID: 15489334
Boster Scientific Support
Answered: 2019-07-01
Question
We have been able to see staining in mouse placenta. Do you have any suggestions? Is anti-Aryl hydrocarbon Receptor/Ahr antibody supposed to stain placenta positively?
E. Zhang
Verified customer
Asked: 2017-06-28
Answer
Based on literature placenta does express AHR. Based on Uniprot.org, AHR is expressed in placenta, liver, among other tissues. Regarding which tissues have AHR expression, here are a few articles citing expression in various tissues:
Liver, Pubmed ID: 8393992
Placenta, Pubmed ID: 15489334
Boster Scientific Support
Answered: 2017-06-28
Question
We ordered your anti-Aryl hydrocarbon Receptor/Ahr antibody for WB on placenta a few months ago. I am using mouse, and We intend to use the antibody for Flow Cytometry next. My question regards examining placenta as well as liver in our next experiment. Do you have any suggestion on which antibody would work the best for Flow Cytometry?
S. Banerjee
Verified customer
Asked: 2013-06-28
Answer
I looked at the website and datasheets of our anti-Aryl hydrocarbon Receptor/Ahr antibody and I see that A00225-3 has been validated on mouse in both WB and Flow Cytometry. Thus A00225-3 should work for your application. Our Boster satisfaction guarantee will cover this product for Flow Cytometry in mouse even if the specific tissue type has not been validated. We do have a comprehensive range of products for Flow Cytometry detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2013-06-28
