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Product Info Summary
| SKU: | P00059 |
|---|---|
| Size: | 100ug |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, IHC, WB |
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Product info
Product Name
Anti-Phospho-SMAD3 S423/S425 Antibody
SKU/Catalog Number
P00059
Size
100ug
Form
Liquid (sterile filtered)
Description
Boster Bio Anti-Phospho-SMAD3 S423/S425 Antibody (Catalog # P00059). Tested in ELISA, IHC, WB applications. This antibody reacts with Human.
Storage & Handling
Store vial at -20°C prior to opening. Aliquot contents and freeze at -20°C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4°C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening. (Ship on dry ice.)
Cite This Product
Anti-Phospho-SMAD3 S423/S425 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # P00059)
Host
Rabbit
Contents
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 0.01% (w/v) Sodium Azide
Clonality
Polyclonal
Isotype
IgG
Immunogen
Anti-SMAD3 pS423pS425 antibody was prepared from whole rabbit serum produced by repeated immunizations with a dual phosphorylated synthetic peptide corresponding to a c-terminal region with Serine 423 and Serine 425 of human SMAD3 protein.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross reactivity with other proteins.
Reactive Species
P00059 is reactive to SMAD3 in Human
Reconstitution
Calculated molecular weight
48081 MW
Background of SMAD3
This antibody is suitable for Cancer, Immunology and Nuclear Signaling research. Smad3 (also known as Mothers against decapentaplegic homolog 3 Mothers against DPP homolog 3, Mad3, hMAD-3, JV15-2 or hSMAD3) is a transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinase. These activators exert diverse effects on a wide array of cellular processes. The Smad proteins mediate much of the signaling responses induced by the TGF-b superfamily. Briefly, activated type I receptor phosphorylates receptor-activated Smads (R-Smads) at their c-terminal two extreme serines in the SSXS motif, e.g. Smad2 and Smad3 proteins in the TGF-b pathway, or Smad1, Smad5 or Smad8 in the BMP pathway. Then the phosphorylated R-Smad translocated into nucleus, where they regulate transcription of target genes. Based on microarray and animal model experiments, Smad3 accounts for at least 80% of all TGF-b-mediated response.
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
P00059 is guaranteed for ELISA, 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.
ELISA: 1:15,000 - 1:30,000
Flow Cytometry: User optimized
IHC: 1:500
WB: 1:2,000 - 1:20,000
Validation Images & Assay Conditions
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SMAD3 was detected in paraffin-embedded sections of epidermal keratinocytes at 40X using rabbit anti-SMAD3 phospho S423/phospho S425 Antigen Affinity purified polyclonal antibody (Catalog # P00059) at 2.5 μg/mL. The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain. Personal Communi-cation, Tina Roush, LifeSpanBiosciences, Seattle, WA.
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Western blot analysis of endogenous SMAD3 expression in stimulated cell lysates. Lysates were prepared from control cells (-lanes), or cells stimulated with 2 ng/ml TGF for 1 hour. Phosphorylated SMAD3 was detected using rabbit anti-SMAD3 phospho S423/phospho S425 Antigen Affinity purified polyclonal antibody (Catalog # P00059). Personal Communication. Ying Zhang, NIH, CCR, Bethesda, MD.
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Western blot analysis of SMAD3 expression in HEK293 lysate (lane 1), HeLa Lysate (lane 2), MCF-7 Lysate (lane 3), Jurkat Lysate (lane 4), A431 Lysate (lane 5), A549 Lysate (lane 6), LNCap Lysate (lane 7), MOLT-4 Lysate (lane 8), Ramos Lysate (lane 9), Raji Lsyate (lane 10), A-172 Lysate (lane 11) and NIH/3T3 Lysate (lane 12). Phosphorylated SMAD3 at 35KD was detected using rabbit anti-SMAD3 phospho S423/phospho S425 Antigen Affinity purified polyclonal antibody (Catalog # P00059) at 1ug/mL. The blot was developed using chemiluminescence (ECL) method (Catalog # EK1002).
Protein Target Info & Infographic
Gene/Protein Information For SMAD3 (Source: Uniprot.org, NCBI)
Gene Name
SMAD3
Full Name
Mothers against decapentaplegic homolog 3
Weight
48081 MW
Superfamily
dwarfin/SMAD family
Alternative Names
SMAD3 pS423pS425, SMAD-3, SMAD 3, mothers against decapentaplegic homolog 3, MAD homolog 3, Mothers against DPP homolog 3, SMAD family member 3, MADH3, MADH 3, JV15-2, nothing SMAD3 HSPC193, HsT17436, JV15-2, LDS1C, LDS3, MADH3 SMAD family member 3 mothers against decapentaplegic homolog 3|MAD homolog 3|MAD, mothers against decapentaplegic homolog 3|SMA- and MAD-related protein 3|SMAD, mothers against DPP homolog 3|hMAD-3|hSMAD3|mad homolog JV15-2|mad protein homolog|mad3|mothers against DPP homolog 3
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on SMAD3, check out the SMAD3 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for SMAD3: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-Phospho-SMAD3 S423/S425 Antibody (P00059)
Hello CJ!
P00059 has been cited in 4 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
Wang Q,Liu J,Hu Y,Pan T,Xu Y,Yu J,Xiong W,Zhou Q,Wang Y.Local administration of liposomal-based Srpx2 gene therapy reverses pulmonary fibrosis by blockading fibroblast-to-myofibroblast transition.Theranostics.2021 May 13;11(14):7110-7125.doi:10.7150/thno. 61085. PMID:34093874;PMCID:PMC8171094.
Species: Human,Mouse
P00059 usage in article: APP:WB, SAMPLE:FIBROBLASTS AND LUNG TISSUE, DILUTION:NA
Hong L,Li F,Tang C,Li L,Sun L,Li X,Zhu L.Semaphorin 7A promotes endothelial to mesenchymal transition through ATF3 mediated TGF-β2/Smad signaling.Cell Death Dis.2020 Aug 10;11(8):695.doi:10.1038/s41419-020-02818-x.PMID:32826874;PMCID:PMC7442651.
Species: Human,Mouse
P00059 usage in article: APP:WB, SAMPLE:HUVECS, DILUTION:NA
Shi K, Jiang J, Ma T, Xie J, Duan L, Chen R, Song P, Yu Z, Liu C, Zhu Q, Zheng J. Int J Clin Exp Med. 2014 Sep 15;7(9):2645-50. Ecollection 2014. Dexamethasone Attenuates Bleomycin-Induced Lung Fibrosis In Mice Through Tgf-??, Smad3 And Jak-Stat P...
Xu T, Ni Mm, Huang C, Meng Xm, He Yh, Zhang L, Li J. Inflammation. 2015 Oct;38(5):1794-804. Doi: 10.1007/S10753-015-0157-6. Nlrc5 Mediates Il-6 And Il-1?? Secretion In Lx-2 Cells And Modulated By The Nf-??b/Smad3 Pathway.
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Customer Reviews
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Customer Q&As
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15 Customer Q&As for Anti-Phospho-SMAD3 S423/S425 Antibody
Question
Does P00059 anti-Phospho-SMAD3 S423/S425 antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2020-03-30
Answer
You can see on the product datasheet, P00059 anti-Phospho-SMAD3 S423/S425 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: 2020-03-30
Question
We want using your anti-Phospho-SMAD3 S423/S425 antibody for negative regulation of inflammatory response studies. Has this antibody been tested with western blotting on or cells? We would like to see some validation images before ordering.
Verified Customer
Verified customer
Asked: 2020-01-29
Answer
We appreciate your inquiry. This P00059 anti-Phospho-SMAD3 S423/S425 antibody is validated on or cells. It is guaranteed to work for IHC, 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: 2020-01-29
Question
Is a blocking peptide available for product anti-Phospho-SMAD3 S423/S425 antibody (P00059)?
Verified Customer
Verified customer
Asked: 2020-01-20
Answer
We do provide the blocking peptide for product anti-Phospho-SMAD3 S423/S425 antibody (P00059). If you would like to place an order for it please contact [email protected] and make a special request.
Boster Scientific Support
Answered: 2020-01-20
Question
Thank you for helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for erythroleukemia using anti-Phospho-SMAD3 S423/S425 antibody P00059. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2020-01-16
Answer
We 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: 2020-01-16
Question
Is there a BSA free version of anti-Phospho-SMAD3 S423/S425 antibody P00059 available?
Verified Customer
Verified customer
Asked: 2020-01-10
Answer
Thanks for your recent telephone inquiry. I can confirm that some lots of this anti-Phospho-SMAD3 S423/S425 antibody P00059 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: 2020-01-10
Question
We were content with the WB result of your anti-Phospho-SMAD3 S423/S425 antibody. However we have observed positive staining in erythroleukemia cytoplasm using this antibody. Is that expected? Could you tell me where is SMAD3 supposed to be expressed?
Verified Customer
Verified customer
Asked: 2020-01-07
Answer
From literature, erythroleukemia 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: 2020-01-07
Question
We are currently using anti-Phospho-SMAD3 S423/S425 antibody P00059 for human tissue, and we are well pleased with the IHC results. The species of reactivity given in the datasheet says human, mouse, rat. Is it true that the antibody can work on horse tissues as well?
Verified Customer
Verified customer
Asked: 2019-07-23
Answer
The anti-Phospho-SMAD3 S423/S425 antibody (P00059) has not been tested for cross reactivity specifically with horse 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 horse you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-07-23
Question
We ordered your anti-Phospho-SMAD3 S423/S425 antibody for WB on erythroleukemia a few months ago. I am using mouse, and We are going to use the antibody for IHC next. you antibody examining erythroleukemia as well as colon carcinoma in our next experiment. Could you please give me some suggestion on which antibody would work the best for IHC?
Verified Customer
Verified customer
Asked: 2019-03-15
Answer
I viewed the website and datasheets of our anti-Phospho-SMAD3 S423/S425 antibody and I see that P00059 has been validated on mouse in both WB and IHC. Thus P00059 should work for your application. Our Boster satisfaction guarantee will cover this product for IHC in mouse even if the specific tissue type has not been validated. We do have a comprehensive range of products for IHC detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2019-03-15
Question
Here is the WB image, lot number and protocol we used for erythroleukemia using anti-Phospho-SMAD3 S423/S425 antibody P00059. Please let me know if you require anything else.
P. Rodriguez
Verified customer
Asked: 2018-09-27
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: 2018-09-27
Question
I was wanting to use your anti-Phospho-SMAD3 S423/S425 antibody for WB for mouse erythroleukemia on frozen tissues, but I want to know if it has been validated for this particular application. Has this antibody been validated and is this antibody a good choice for mouse erythroleukemia identification?
Verified Customer
Verified customer
Asked: 2018-03-20
Answer
As indicated on the product datasheet, P00059 anti-Phospho-SMAD3 S423/S425 antibody has been validated for IHC, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in mouse erythroleukemia in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2018-03-20
Question
We have seen staining in rat erythroleukemia. Do you have any suggestions? Is anti-Phospho-SMAD3 S423/S425 antibody supposed to stain erythroleukemia positively?
Verified Customer
Verified customer
Asked: 2018-03-02
Answer
From literature erythroleukemia does express SMAD3. From 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: 2018-03-02
Question
I see that the anti-Phospho-SMAD3 S423/S425 antibody P00059 works with WB, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2017-08-23
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 [email protected]
Boster Scientific Support
Answered: 2017-08-23
Question
I would like to test anti-Phospho-SMAD3 S423/S425 antibody P00059 on mouse erythroleukemia for research purposes, then I may be interested in using anti-Phospho-SMAD3 S423/S425 antibody P00059 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
Verified Customer
Verified customer
Asked: 2017-08-11
Answer
The products we sell, including anti-Phospho-SMAD3 S423/S425 antibody P00059, 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: 2017-08-11
Question
I have a question about product P00059, anti-Phospho-SMAD3 S423/S425 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?
G. Dhar
Verified customer
Asked: 2017-01-16
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 P00059 anti-Phospho-SMAD3 S423/S425 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: 2017-01-16
Question
Would anti-Phospho-SMAD3 S423/S425 antibody P00059 work for WB with erythroleukemia?
J. Krishna
Verified customer
Asked: 2016-02-16
Answer
According to the expression profile of erythroleukemia, SMAD3 is highly expressed in erythroleukemia. So, it is likely that anti-Phospho-SMAD3 S423/S425 antibody P00059 will work for WB with erythroleukemia.
Boster Scientific Support
Answered: 2016-02-16
