Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] (ab214423)
![Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] (ab214423)](https://www.abcam.com/ps/products/214/ab214423/Images/ab214423-284412-anti-smad1-phospho-s463-s465-antibody-epr20662-29-western-blot.jpg "Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] (ab214423)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20662-29] to Smad1 (phospho S463 + S465)
- Suitable for: WB, Dot blot, IP
- Reacts with: Mouse, Human
Overview
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Product name
Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29]
See all Smad1 primary antibodies -
Description
Rabbit monoclonal [EPR20662-29] to Smad1 (phospho S463 + S465) -
Host species
Rabbit -
Specificity
Based on sequence homology this antibody also reacts with Smad5 (phospho S463/S465) and Smad9 (phospho S465/S467). -
Tested Applications & Species
See all applications and species dataApplication Species IP MouseWB MouseHuman
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Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Dot Blot: Smad1 (phospho S463/S465) peptide, Smad1 (phospho S465) peptide, Smad5 (phospho S463/S465) peptide, Smad5 (phospho S465) peptide. WB: Calyculin (ab141784) treated HeLa cell lysate; BMP2 treated NIH/3T3 cell lysate. IP: BMP2 treated NIH/3T3 cell lysate.
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General notes
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20662-29 -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] (ab214423)All lanes : Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] (ab214423) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) grown in serum-free media overnight, whole cell lysate
Lane 2 : HeLa grown in serum-free media overnight, then treated with 100ng/ml Calyculin A (ab141784) for 15 minutes, Calyculin A was removed, followed by treatment with 100ng/ml BMP2 for 30 minutes, whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast) grown in serum-free media overnight, whole cell lysate
Lane 4 : NIH/3T3 grown in serum-free media overnight, then treated with 50ng/ml BMP2 for 30 minutes, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 52 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking/Dilution: 5% NFDM/TBST.
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![Dot Blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] (ab214423)](https://www.abcam.com/ps/products/214/ab214423/Images/ab214423-284409-anti-smad1-phospho-s463-s465-antibody-epr20662-29-dot-blot.jpg)
Dot Blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] (ab214423)Dot blot analysis of Smad1 (phospho S463/S465) peptide (Lane 1), Smad1 (phospho S463) peptide (Lane 2), Smad1 (phospho S465) peptide (Lane 3), Smad1 non-phospho peptide (Lane 4), Smad5 (phospho S463/S465) peptide (Lane 5), Smad5 (phospho S463) peptide (Lane 6), Smad5 (phospho S465) peptide (Lane 7) and Smad5 non-phospho peptide (Lane 8) using ab214423 at 1/1,000 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100,000 dilution.
Blocking and Diluting buffer and concentration: 5% NFDM /TBST.
Exposure time: 30 seconds.
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![Immunoprecipitation - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] (ab214423)](https://www.abcam.com/ps/products/214/ab214423/Images/ab214423-284422-anti-smad1-phospho-s463-s465-antibody-epr20662-29-immunoprecipitation.jpg)
Immunoprecipitation - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] (ab214423)Smad1 (phospho S463/S465) was immunoprecipitated from 0.35 mg of NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate with ab214423 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab214423 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution
Lane 1: NIH/3T3 (mouse embryonic fibroblast) grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate 10 μg (Input).
Lane 2: ab214423 IP in NIH/3T3 grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214423 in NIH/3T3 grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysateBlocking and dilution buffer: 5% NFDM/TBST.
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![Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] (ab214423)](https://www.abcam.com/ps/products/214/ab214423/Images/ab214423-4-benefits-of-recombinant-antibodies.png)
Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] (ab214423)
