Overview

  • Product name
    Anti-SMAD1/3/5 (phospho S423 + S425) antibody
  • Description
    Rabbit polyclonal to SMAD1/3/5 (phospho S423 + S425)
  • Host species
    Rabbit
  • Specificity
    ab51451 is specific for the phosphorylated form of the protein although minimal reactivity with nonphosphorylated human Smad3 can be observed by ELISA and WB. ab51451 also reacts with phosphorylated Smad1 and Smad5 but negligible reactivity is seen against other phosphorylated Smad family members.
  • Tested applications
    Suitable for: IHC-P, WB, ELISA, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Chicken, Human
    Predicted to work with: Rat, Cow, Pig, Xenopus laevis, Zebrafish
  • Immunogen

    Synthetic peptide corresponding to Human SMAD1/3/5 aa 417-425.
    Sequence:

    PSIRCSSVS

Properties

Applications

Our Abpromise guarantee covers the use of ab51451 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use at an assay dependent concentration. PubMed: 20519446
WB 1/2000 - 1/20000. Predicted molecular weight: 48 kDa. Stimulation with 2 ng/ml TGF-beta for 1 hour is suggested.
ELISA 1/25000 - 1/100000.
ICC/IF Use at an assay dependent concentration.

Target

Images

  • All lanes : Anti-SMAD1/3/5 (phospho S423 + S425) antibody (ab51451) at 1 µg/ml

    Lane 1 : HEK293 lysate
    Lane 2 : HeLa Lysate
    Lane 3 : MCF-7 Lysate
    Lane 4 : Jurkat Lysate
    Lane 5 : A431 Lysate
    Lane 6 : A549 Lysate
    Lane 7 : LNCap Lysate
    Lane 8 : MOLT-4 Lysate
    Lane 9 : Ramos Lysate
    Lane 10 : Raji Lysate
    Lane 11 : A-172 Lysate
    Lane 12 : NIH/3T3 Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lanes 1-2 & 4-12 : Peroxidase rabbit secondary antibody at 1/30000 dilution
    Lane 3 : Peroxidase rabbit secondary antibody

    Predicted band size: 48 kDa

  • ab51451 at 2.5 µg/ml staining epidermal keratinocytes at 40X.  This image shows strong nuclear staining with localization of the antibody as the precipated red siganl and a hematoxylin purple nuclear counterstain.

  • ab51451 staining Smad3 (phospho S423 + S425) in PC3 and LnCAP cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 70% ethanol and blocked for 1 hour at 26°C. Samples were incubated with primary antibody (1/150) for 16 hours at 4°C. An IRDye® 800CW-conjugated goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody. NC-PC3 - negative control PC3 cells, NC-LnCAP - negative control LnCAP cells.

    See Abreview

  • All lanes : Anti-SMAD1/3/5 (phospho S423 + S425) antibody (ab51451) at 1/2000 dilution

    Lane 1 : MEF lysate (control)
    Lane 2 : MEF lysate, after stimulation with 2 ng/ml of TGF-ß for 1 hour
    Lane 3 : MEF-/-Smad3 lysate (control)
    Lane 4 : MEF-/-Smad3 lysate, after stimulation with 2 ng/ml of TGF-ß for one hour
    Lane 5 : Hep3B lysate (control)
    Lane 6 : Hep3B lysate, after stimulation with 2 ng/ml of TGF-ß for one hour
    Lane 7 : HEK 293 lysate (control)

    Predicted band size: 48 kDa



    Image courtesy of Ying Zhang, NIH, CCR, Bethesda, MD.

References

This product has been referenced in:
  • Murugavel S  et al. Valproic Acid Induces Endothelial-to-Mesenchymal Transition-Like Phenotypic Switching. Front Pharmacol 9:737 (2018). Read more (PubMed: 30050438) »
  • Quattrocelli M  et al. Genetic modifiers of muscular dystrophy act on sarcolemmal resealing and recovery from injury. PLoS Genet 13:e1007070 (2017). Read more (PubMed: 29065150) »
See all 35 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Western blot
Sample
Rat Tissue lysate - whole (Muscle)
Gel Running Conditions
Reduced Denaturing (9)
Loading amount
50 µg
Specification
Muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Mr. Baptiste Jude

Verified customer

Submitted Feb 26 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (heart)
Permeabilization
Yes - 0.3% Triton X
Specification
heart
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 20% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 19 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C
Sample
Human Cell (PC3 and LnCAP)
Specification
PC3 and LnCAP
Permeabilization
Yes - 70% Ethanol
Fixative
Paraformaldehyde

Dr. Francesco Elia Marino

Verified customer

Submitted May 02 2014

Application
Western blot
Loading amount
60 µg
Gel Running Conditions
Reduced Non-Denaturing (Native) (12)
Sample
Mouse Tissue lysate - whole (Mouse gastrocnemius)
Specification
Mouse gastrocnemius
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C

Dr. Francesco Elia Marino

Verified customer

Submitted Mar 26 2014

Question
Answer

Thank you for calling Abcam earlier today.

As we talked about on the telephone, I am sending you ab52903 as a free of charge replacement. Your new order/reference number is********.

Please let me know if there is anything else I can help you with.

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Answer

Thank you for your reply.

The only other phospho-smad3 (S423+S425) we have are:

ab11825 - https://www.abcam.com/Smad3-phospho-S423-S425-antibody-ab118825.html

ab52903 - https://www.abcam.com/Smad3-phospho-S423-S425-antibody-EP823Y-ab52903.html



Please let me know which one you would like to test and I will then send you the testing discount codes for it.

Also for the other antibodies, below are some selections:

PRKACA,

ab124390 - https://www.abcam.com/PRKACA-antibody-X1-ab124390.html

Stat3
ab119352 - https://www.abcam.com/STAT3-antibody-9D8-ab119352.html

phospho-Stat3
ab30647 - https://www.abcam.com/STAT3-phospho-S727-antibody-ab30647.html
ab30646 - https://www.abcam.com/STAT3-phospho-Y705-antibody-ab30646.html



Above are only a few of the antibodies that we have for these targets, so please feel free to search and see if you can find a more suitable alternative. Also non have been tested in flow cytometry and so would qualify for a testing discount code.

Please let me know if there is anything else I can help you with.

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Answer

Thank you very much for contacting Abcam.

The isotype control that I would recommend for your flow cytometry experiments is:

https://www.abcam.com/Rabbit-IgG-ChIP-Grade-ab37415.html



Below is the infromation about the testing discount program, you would be eligbale to get a testing disount for all 3 antibodies:

https://www.abcam.com/TGF-beta-Receptor-I-antibody-ab125310.html
https://www.abcam.com/Smad3-phospho-S423-S425-antibody-ab51451.html
https://www.abcam.com/Smad3-antibody-ChIP-Grade-ab28379.html



To our knowledge, ab125310/ab28379/ab51451 has not been tested in Flow Cytometry. Therefore, I can offer a discount off a future purchase if you buy ab125310/ab28379/ab51451 now, test it in Flow Cytometry and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free PRIMARY ANTIBODY for each antibody that is tested in Flow Cytometry.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab125310/ab28379/ab51451 in Flow Cytometry. I will then send a discount code. This code must be issued before purchasing ab125310/ab28379/ab51451 so please wait for my reply before ordering.

2. Purchase ab125310/ab28379/ab51451 either by phone, fax, or online (www.abcam.com).

3. Test it in Flow Cytometry.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any each PRIMARY ANTIBODY ordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab125310/ab28379/ab51451 turns out to be unsuitable for Flow Cytometry, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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