Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19686] to Smad3 - BSA and Azide free
  • Suitable for: ChIP, WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Smad3 antibody [EPR19686] - BSA and Azide free
    See all Smad3 primary antibodies

  • Description

    Rabbit monoclonal [EPR19686] to Smad3 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ChIP, WB, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A549 and HeLa cell lysates; Human kidney cell lysate. ChIP: Chromatin was prepared from HaCaT cells.

  • General notes

    Ab251490 is the carrier-free version of ab208182. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab251490 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Images

  • Western blot - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490)
    Western blot - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490)
    All lanes : Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182) at 1/1000 dilution

    Lane 1 : Wild-type A549 cell lysate
    Lane 2 : SMAD3 knockout A549 cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : Human Kidney cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 48 kDa
    Observed band size: 50 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab208182).

    Lanes 1 - 4: Merged signal (red and green). Green - ab208182 observed at 50 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab208182 was shown to react with Smad3 in western blot. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab208182 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • ChIP - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490)
    ChIP - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490)

    This data was developed using the same antibody clone in a different buffer formulation (ab208182).

    Chromatin was prepared from HaCaT (Human keratinocyte cell line) cells treated with 7ng/ml TGF-β for 1h and non-treated according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab208182 (red), and 20µl of protein A/G beads. 2μg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

    The ChIP condition performed here is similar to the literature (PMID: 18245174).

  • Western blot - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490)
    Western blot - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490)
    All lanes : Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : SMAD3 OG knockout HeLa cell lysate
    Lane 3 : HeLa wildtype cell lysate
    Lane 4 : SMAD3 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 48 kDa
    Observed band size: 74 kDa why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab208182).

    Lanes 1-4: Merged signal (red and green). Green - ab208182 observed at 74 kDa. Red - loading control, ab8245 observed at 37 kDa.

    ab208182 was shown to react with Smad3 in wildtype HeLa. Loss of signal was observed when knockout HeLa sample ab263834 was used. Wild-type and Smad3 knockout samples were subjected to SDS-PAGE. ab208182 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)  and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490)
    Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490)

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