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Product Info Summary
| SKU: | PB9396 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Smad3 Antibody Picoband®
SKU/Catalog Number
PB9396
PB0445 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Smad3 Antibody Picoband® catalog # PB9396. Tested in ICC/IF, 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-Smad3 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9396)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence in the middle region of human Smad3, identical to the related mouse and rat sequences.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
PB9396 is reactive to SMAD3 in Human, Mouse, Rat
Observed Molecular Weight
50-55 kDa
Calculated molecular weight
48.1 kDa
Background of SMAD3
Mothers against decapentaplegic homolog 3 also known as SMAD family member 3 or SMAD3 is a protein that in humans is encoded by the SMAD3 gene. SMAD3 is a member of the SMAD family of proteins. It is located in 15q22.33. And it mediates the signals from the transforming growth factor beta (TGF-β) super family ligands that regulate cell proliferation, differentiation and death. Based on its essential role in TGF beta signaling pathway, SMAD3 has been related with tumor growth in cancer development.
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
PB9396 is guaranteed for 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, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Positive Control
WB: human A549 whole cell, human RT4 whole cell, human A431 whole cell, human PC-3 whole cell, rat brain tissue, rat ovary tissue, mouse brain tissue
IHC: human liver cancer tissuebr>ICC/IF: A549 cell
Validation Images & Assay Conditions
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Western blot analysis of Smad3 using anti-Smad3 antibody (PB9396).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human A549 whole cell lysates,
Lane 2: human RT4 whole cell lysates,
Lane 3: human A431 whole cell lysates,
Lane 4: human PC-3 whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat ovary tissue lysates,
Lane 7: 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-Smad3 antigen affinity purified polyclonal antibody (PB9396) 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 (Catalog # BA1054) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for Smad3 at approximately 50-55 kDa. The expected band size for Smad3 is at 48 kDa.
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IHC analysis of Smad3 using anti-Smad3 antibody (PB9396).
Smad3 was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Smad3 Antibody (PB9396) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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IF analysis of Smad3 using anti-Smad3 antibody (PB9396) and anti-Tubulin Alpha antibody (M03989-3).
Smad3 was detected in immunocytochemical section of A549 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-Smad3 Antibody (PB9396) and mouse anti-Tubulin Alpha antibody (M03989-3) overnight at 4°C. FITC Conjugated Goat Anti-Rabbit IgG (BA1105) and Cy3 Conjugated Goat Anti-Mouse IgG (BA1031) 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.
Specific Publications For Anti-Smad3 Antibody Picoband® (PB9396)
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Customer Q&As
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15 Customer Q&As for Anti-Smad3 Antibody Picoband®
Question
Would anti-Smad3 antibody PB9396 work for WB with pancreas?
Verified Customer
Verified customer
Asked: 2020-04-27
Answer
According to the expression profile of pancreas, SMAD3 is highly expressed in pancreas. So, it is likely that anti-Smad3 antibody PB9396 will work for WB with pancreas.
Boster Scientific Support
Answered: 2020-04-27
Question
Do you have a BSA free version of anti-Smad3 antibody PB9396 available?
Verified Customer
Verified customer
Asked: 2020-03-11
Answer
Thanks for your recent telephone inquiry. I can confirm that some lots of this anti-Smad3 antibody PB9396 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: 2020-03-11
Question
I was wanting to use your anti-Smad3 antibody for WB for human pancreas 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 pancreas identification?
Verified Customer
Verified customer
Asked: 2019-10-29
Answer
You can see on the product datasheet, PB9396 anti-Smad3 antibody has been tested for WB on human tissues. We have an innovator award program that if you test this antibody and show it works in human pancreas in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2019-10-29
Question
Will PB9396 anti-Smad3 antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2019-08-16
Answer
It shows on the product datasheet, PB9396 anti-Smad3 antibody as been validated 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: 2019-08-16
Question
Our lab were satisfied with the WB result of your anti-Smad3 antibody. However we have seen positive staining in amniotic fluid cytoplasm using this antibody. Is that expected? Could you tell me where is SMAD3 supposed to be expressed?
Verified Customer
Verified customer
Asked: 2019-08-15
Answer
From literature, amniotic fluid does express SMAD3. Generally SMAD3 expresses in cytoplasm. Regarding which tissues have SMAD3 expression, here are a few articles citing expression in various tissues:
Brain, Pubmed ID: 14702039
Cervix carcinoma, Pubmed ID: 18669648
Colon carcinoma, Pubmed ID: 9464505
Erythroleukemia, Pubmed ID: 23186163
Pancreas, Pubmed ID: 15489334
Placenta, Pubmed ID: 8774881
Boster Scientific Support
Answered: 2019-08-15
Question
My question regards to test anti-Smad3 antibody PB9396 on human pancreas for research purposes, then I may be interested in using anti-Smad3 antibody PB9396 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
Verified Customer
Verified customer
Asked: 2019-07-30
Answer
The products we sell, including anti-Smad3 antibody PB9396, 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: 2019-07-30
Question
Thank you for helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for pancreas using anti-Smad3 antibody PB9396. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2019-04-08
Answer
I appreciate 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: 2019-04-08
Question
I have a question about product PB9396, anti-Smad3 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: 2018-12-12
Answer
We do not recommend 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 PB9396 anti-Smad3 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: 2018-12-12
Question
We are currently using anti-Smad3 antibody PB9396 for human tissue, and we are well pleased with the WB results. The species of reactivity given in the datasheet says human. Is it possible that the antibody can work on feline tissues as well?
Verified Customer
Verified customer
Asked: 2018-10-22
Answer
The anti-Smad3 antibody (PB9396) has not been tested for cross reactivity specifically with feline tissues, but 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: 2018-10-22
Question
Please see the WB image, lot number and protocol we used for pancreas using anti-Smad3 antibody PB9396. Please let me know if you require anything else.
Verified Customer
Verified customer
Asked: 2018-08-03
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: 2018-08-03
Question
Is this PB9396 anti-Smad3 antibody reactive to the isotypes of SMAD3?
Verified Customer
Verified customer
Asked: 2018-01-12
Answer
The immunogen of PB9396 anti-Smad3 antibody is A synthetic peptide corresponding to a sequence in the middle region of human Smad3 (193-222aa DHQMNHSMDAGSPNLSPNPMSPAHNNLDLQ), identical to the related mouse and rat sequences. 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: 2018-01-12
Question
Is a blocking peptide available for product anti-Smad3 antibody (PB9396)?
Verified Customer
Verified customer
Asked: 2017-06-02
Answer
We do provide the blocking peptide for product anti-Smad3 antibody (PB9396). If you would like to place an order for it please contact support@bosterbio.com and make a special request.
Boster Scientific Support
Answered: 2017-06-02
Question
I see that the anti-Smad3 antibody PB9396 works with WB, what is the protocol used to produce the result images on the product page?
H. Miller
Verified customer
Asked: 2015-05-06
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: 2015-05-06
Question
I am looking for using your anti-Smad3 antibody for smad protein signal transduction studies. Has this antibody been tested with western blotting on u87 whole cell lysate? We would like to see some validation images before ordering.
M. Thomas
Verified customer
Asked: 2014-12-05
Answer
I appreciate your inquiry. This PB9396 anti-Smad3 antibody is tested on u87 whole cell lysate, human placenta tissue, tissue lysate, panc whole cell lysate, hela whole cell lysate. It is guaranteed to work for WB in human. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2014-12-05
Question
We have observed staining in human amniotic fluid. Any tips? Is anti-Smad3 antibody supposed to stain amniotic fluid positively?
H. Gonzalez
Verified customer
Asked: 2013-01-17
Answer
From what I have seen in literature amniotic fluid does express SMAD3. From what I have seen in Uniprot.org, SMAD3 is expressed in amniotic fluid, placenta, colon carcinoma, brain, pancreas, cervix carcinoma, erythroleukemia, among other tissues. Regarding which tissues have SMAD3 expression, here are a few articles citing expression in various tissues:
Brain, Pubmed ID: 14702039
Cervix carcinoma, Pubmed ID: 18669648
Colon carcinoma, Pubmed ID: 9464505
Erythroleukemia, Pubmed ID: 23186163
Pancreas, Pubmed ID: 15489334
Placenta, Pubmed ID: 8774881
Boster Scientific Support
Answered: 2013-01-17
