Product Info Summary
| SKU: | PB9474 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-Aquaporin 2/AQP2 Antibody Picoband®
SKU/Catalog Number
PB9474
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Aquaporin 2/AQP2 Antibody Picoband® catalog # PB9474. Tested in Flow Cytometry, IHC, 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
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-Aquaporin 2/AQP2 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9474)
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
A synthetic peptide corresponding to a sequence at the C-terminus of human Aquaporin 2, different from the related mouse and rat sequences by one amino acid.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9474 is reactive to AQP2 in Human, Mouse, Rat
Observed Molecular Weight
26 kDa
Calculated molecular weight
28.8 kDa
Background of AQP2
AQP2 (Aquaporin 2), also called AQUAPORIN-CD, is found in the apical cell membranes of the kidney's collecting duct principal cells and in intracellular vesicles located throughout the cell. The AQP2 gene is mapped to chromosome 12q13, very close to the site of major intrinsic protein by situ hybridization. The investigators suggested that a defect in the AQP2 gene is the basis of the autosomal form of nephrogenic diabetes insipidus. The functional expression and the limited localization suggested that AQP2 is the vasopressin-regulated water channel. Using rat kidney slices and porcine kidney cells stably expressing rat Aqp2, AQP2 trafficking can be stimulated by cAMP-independent pathways that utilize nitric oxide (NO). The NO donors sodium nitroprusside (SNP) and NONOate and the NO synthase substrate L-arginine mimicked the effect of vasopressin (VP), stimulating relocation of Aqp2 from cytoplasmic vesicles to the apical plasma membrane. SNP increased intracellular cGMP rather than cAMP, and exogenous cGMP stimulated AQP2 membrane insertion. Atrial natriuretic factor, which signals via cGMP, also stimulated AQP2 translocation. AQP2 expression in kidney connecting tubules is sufficient for survival and that AQP2 expression in collecting ducts is required to regulate body water balance. The S256L substitution in the cytoplasmic tail of the Aqp2 protein prevented phosphorylation at S256 and the subsequent accumulation of Aqp2 on the apical membrane of the collecting duct principal cells.
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
PB9474 is guaranteed for 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, 0.1-0.5μg/ml, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: rat kidney tissue, rat NRK whole cell, rat PC-12 whole cell, mouse kidney tissue, mouse HBZY whole cell, mouse RAW2647 whole cell
IHC: Mouse Kidney tissue, Rat Kidney tissue, Human Renal Cancer tissue
FCM: PC-3 cell
Validation Images & Assay Conditions
Click image to see more details
Western blot analysis of Aquaporin 2 using anti-Aquaporin 2 antibody (PB9474).
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: rat kidney tissue lysates,
Lane 2: rat NRK whole cell lysates,
Lane 3: rat PC-12 whole cell lysates,
Lane 4: mouse kidney tissue lysates,
Lane 5: mouse HBZY whole cell lysates,
Lane 6: mouse RAW264.7 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-Aquaporin 2 antigen affinity purified polyclonal antibody (Catalog # PB9474) 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 Aquaporin 2 at approximately 26 kDa. The expected band size for Aquaporin 2 is at 26 kDa.
Click image to see more details
IHC analysis of Aquaporin 2 using anti-Aquaporin 2 antibody (PB9474).
Aquaporin 2 was detected in paraffin-embedded section of Mouse Kidney 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-Aquaporin 2 Antibody (PB9474) 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.
Click image to see more details
IHC analysis of Aquaporin 2 using anti-Aquaporin 2 antibody (PB9474).
Aquaporin 2 was detected in paraffin-embedded section of Rat Kidney 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-Aquaporin 2 Antibody (PB9474) 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.
Click image to see more details
IHC analysis of Aquaporin 2 using anti-Aquaporin 2 antibody (PB9474).
Aquaporin 2 was detected in paraffin-embedded section of Human Renal 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-Aquaporin 2 Antibody (PB9474) 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.
Click image to see more details
Flow Cytometry analysis of PC-3 cells using anti-AQP2 antibody (PB9474).
Overlay histogram showing PC-3 cells stained with PB9474 (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-AQP2 Antibody (PB9474,1μg/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-Aquaporin 2/AQP2 Antibody Picoband® (PB9474)
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16 Customer Q&As for Anti-Aquaporin 2/AQP2 Antibody Picoband®
Question
Is a blocking peptide available for product anti-Aquaporin 2/AQP2 antibody (PB9474)?
G. Johnson
Verified customer
Asked: 2020-05-04
Answer
We do provide the blocking peptide for product anti-Aquaporin 2/AQP2 antibody (PB9474). If you would like to place an order for it please contact support@bosterbio.com and make a special request.
Boster Scientific Support
Answered: 2020-05-04
Question
We are currently using anti-Aquaporin 2/AQP2 antibody PB9474 for rat tissue, and we are happy with the WB results. The species of reactivity given in the datasheet says human, mouse, rat. Is it likely that the antibody can work on dog tissues as well?
Verified Customer
Verified customer
Asked: 2020-04-23
Answer
The anti-Aquaporin 2/AQP2 antibody (PB9474) has not been validated for cross reactivity specifically with dog 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 dog you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2020-04-23
Question
I was wanting to use your anti-Aquaporin 2/AQP2 antibody for WB for human cortex of kidney on frozen tissues, but I want to know if it has been tested for this particular application. Has this antibody been tested and is this antibody a good choice for human cortex of kidney identification?
Verified Customer
Verified customer
Asked: 2020-04-21
Answer
As indicated on the product datasheet, PB9474 anti-Aquaporin 2/AQP2 antibody has been validated for Flow Cytometry, IHC, ICC, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in human cortex of kidney in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2020-04-21
Question
Thanks for helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for cortex of kidney using anti-Aquaporin 2/AQP2 antibody PB9474. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2019-11-19
Answer
Thank you for the data. You have provided everything we needed. Our lab team are working to resolve your inquiry as quickly as possible, and we appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2019-11-19
Question
I have attached the WB image, lot number and protocol we used for cortex of kidney using anti-Aquaporin 2/AQP2 antibody PB9474. Please let me know if you require anything else.
Verified Customer
Verified customer
Asked: 2019-06-20
Answer
Thank you very much for the data. Our lab team are working to resolve this as quickly as possible, and we appreciate your patience and understanding! You have provided everything we needed. Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2019-06-20
Question
We have been able to see staining in mouse colon. Are there any suggestions? Is anti-Aquaporin 2/AQP2 antibody supposed to stain colon positively?
L. Jha
Verified customer
Asked: 2019-06-19
Answer
From literature colon does express AQP2. From Uniprot.org, AQP2 is expressed in cortex of kidney, kidney, colon, among other tissues. Regarding which tissues have AQP2 expression, here are a few articles citing expression in various tissues:
Colon, Pubmed ID: 15489334
Kidney, Pubmed ID: 7510718, 11929850
Boster Scientific Support
Answered: 2019-06-19
Question
Would PB9474 anti-Aquaporin 2/AQP2 antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2019-05-31
Answer
As indicated on the product datasheet, PB9474 anti-Aquaporin 2/AQP2 antibody as been tested on WB. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2019-05-31
Question
Will anti-Aquaporin 2/AQP2 antibody PB9474 work for WB with cortex of kidney?
K. Edwards
Verified customer
Asked: 2019-02-27
Answer
According to the expression profile of cortex of kidney, AQP2 is highly expressed in cortex of kidney. So, it is likely that anti-Aquaporin 2/AQP2 antibody PB9474 will work for WB with cortex of kidney.
Boster Scientific Support
Answered: 2019-02-27
Question
Is there a BSA free version of anti-Aquaporin 2/AQP2 antibody PB9474 available?
Verified Customer
Verified customer
Asked: 2018-10-26
Answer
Thanks for your recent telephone inquiry. I can confirm that some lots of this anti-Aquaporin 2/AQP2 antibody PB9474 are BSA free. For now, these lots are available and we can make a BSA free formula for you free of charge. It will take 3 extra days to prepare. If you require this antibody BSA free again in future, please do not hesitate to contact me and I will be pleased to check which lots we have in stock that are BSA free.
Boster Scientific Support
Answered: 2018-10-26
Question
Our lab were well pleased with the WB result of your anti-Aquaporin 2/AQP2 antibody. However we have seen positive staining in cortex of kidney apical cell membrane using this antibody. Is that expected? Could you tell me where is AQP2 supposed to be expressed?
R. Wu
Verified customer
Asked: 2018-09-27
Answer
From literature, cortex of kidney does express AQP2. Generally AQP2 expresses in apical cell membrane. Regarding which tissues have AQP2 expression, here are a few articles citing expression in various tissues:
Colon, Pubmed ID: 15489334
Kidney, Pubmed ID: 7510718, 11929850
Boster Scientific Support
Answered: 2018-09-27
Question
We bought anti-Aquaporin 2/AQP2 antibody for WB on colon in a previous project. I am using human, and We are going to use the antibody for Flow Cytometry next. you antibody examining colon as well as cortex of kidney in our next experiment. Could you please give me some suggestion on which antibody would work the best for Flow Cytometry?
Verified Customer
Verified customer
Asked: 2018-07-09
Answer
I looked at the website and datasheets of our anti-Aquaporin 2/AQP2 antibody and I see that PB9474 has been tested on human in both WB and Flow Cytometry. Thus PB9474 should work for your application. Our Boster satisfaction guarantee will cover this product for Flow Cytometry in human 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: 2018-07-09
Question
Can you help my question with product PB9474, anti-Aquaporin 2/AQP2 antibody. I was wondering if it would be possible to conjugate this antibody with biotin. I would need it to be without BSA or sodium azide. I am planning on using a buffer exchange of sodium azide with PBS only. Would there be problems for me to conjugate the antibody and store it in -20 degrees in small aliquots?
Verified Customer
Verified customer
Asked: 2018-06-21
Answer
We do not advise storing this antibody with PBS buffer only in -20 degrees. If you want to store it in -20 degrees it is best to add some cryoprotectant like glycerol. If you want carrier free PB9474 anti-Aquaporin 2/AQP2 antibody, we can provide it to you in a special formula with trehalose and/or glycerol. These molecules will not interfere with conjugation chemistry and provide a good level of protection for the antibody from degradation. Please be sure to specify this in your purchase order.
Boster Scientific Support
Answered: 2018-06-21
Question
We want using your anti-Aquaporin 2/AQP2 antibody for protein homotetramerization studies. Has this antibody been tested with western blotting on mouse kidney tissue? We would like to see some validation images before ordering.
Verified Customer
Verified customer
Asked: 2018-03-16
Answer
We appreciate your inquiry. This PB9474 anti-Aquaporin 2/AQP2 antibody is tested on rat kidney tissue, tissue lysate, mouse kidney tissue, hela whole cell lysate, a549 whole cell lysate, panc whole cell lysate, renal cancer tissue. It is guaranteed to work for Flow Cytometry, IHC, ICC, WB in human, mouse, rat. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2018-03-16
Question
I am interested in to test anti-Aquaporin 2/AQP2 antibody PB9474 on human cortex of kidney for research purposes, then I may be interested in using anti-Aquaporin 2/AQP2 antibody PB9474 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
L. Krishna
Verified customer
Asked: 2016-11-08
Answer
The products we sell, including anti-Aquaporin 2/AQP2 antibody PB9474, are only intended for research use. They would not be suitable for use in diagnostic work. If you have the means to develop a product into diagnostic use, and are interested in collaborating with us and develop our product into an IVD product, please contact us for more discussions.
Boster Scientific Support
Answered: 2016-11-08
Question
I see that the anti-Aquaporin 2/AQP2 antibody PB9474 works with WB, what is the protocol used to produce the result images on the product page?
Z. Johnson
Verified customer
Asked: 2016-01-22
Answer
You can find protocols for WB on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to support@bosterbio.com
Boster Scientific Support
Answered: 2016-01-22
Question
Is this PB9474 anti-Aquaporin 2/AQP2 antibody reactive to the isotypes of AQP2?
F. Edwards
Verified customer
Asked: 2015-08-07
Answer
The immunogen of PB9474 anti-Aquaporin 2/AQP2 antibody is A synthetic peptide corresponding to a sequence at the C-terminus of human Aquaporin 2 (241-271aa EPDTDWEEREVRRRQSVELHSPQSLPRGTKA), different from the related mouse and rat sequences by one amino acid. Could you tell me which isotype you are interested in so I can help see if the immunogen is part of this isotype?
Boster Scientific Support
Answered: 2015-08-07


