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Recombinant

Recombinant Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)

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Western blot - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
  • Western blot - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
  • Western blot - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
  • Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Mouse monoclonal [18/Smad2/3] to Smad2 + Smad3 - BSA and Azide free
  • Suitable for: IHC-P, WB
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free

  • Description

    Mouse monoclonal [18/Smad2/3] to Smad2 + Smad3 - BSA and Azide free

  • Host species

    Mouse

  • Tested applications

    Suitable for: IHC-P, WBmore details
    Unsuitable for: ICC/IF or IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: His-tagged human Smad2 recombinant protein fragment. His/GST-tagged human Smad3 recombinant protein fragment. HeLa, A549, HT-1080, RAW264.7, C2C12 and C6 whole cell lysate. IHC-P: Human colon and colon cancer tissue. Mouse colon tissue. Mouse breast cancer tissue.

  • General notes

    ab305326 is the carrier-free version of ab305325

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. 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.

Properties

Associated products

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab305326 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. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
Use at an assay dependent concentration. Predicted molecular weight: 48 kDa.
Notes
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
Use at an assay dependent concentration. Predicted molecular weight: 48 kDa.
Application notes
Is unsuitable for ICC/IF or IP.

Target

Images

  • Western blot - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
    Western blot - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
    All lanes : Anti-Smad2 + Smad3 antibody [18/Smad2/3] (ab305325) at 1/1000 dilution

    Lane 1 : His-tagged human Smad2 recombinant protein fragment 10ng
    Lane 2 : His/GST-tagged human Smad3 recombinant protein fragment 10ng

    Secondary
    All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

    Predicted band size: 48 kDa
    Observed band size: 37,60 kDa why is the actual band size different from the predicted?



    This data was developed using ab305325, the same antibody clone in a different buffer formulation.

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Exposure time: 15 seconds.

  • Western blot - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
    Western blot - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
    All lanes : Anti-Smad2 + Smad3 antibody [18/Smad2/3] (ab305325) at 1/1000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate 20 µg
    Lane 2 : A549 (human lung carcinoma epithelial cell), whole cell lysate 20 µg
    Lane 3 : HT-1080 (human fibrosarcoma epithelial cell), whole cell lysate 20 µg

    Secondary
    All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

    Predicted band size: 48 kDa
    Observed band size: 58-62 kDa why is the actual band size different from the predicted?



    This data was developed using ab305325, the same antibody clone in a different buffer formulation. 

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 18809571).

    The identity of the band around 37 kDa is unknown.

    Exposure times: Lanes 1, 4 and 5: 180 seconds; Lanes 2 and 3: 37 seconds.

  • Western blot - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
    Western blot - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
    All lanes : Anti-Smad2 + Smad3 antibody [18/Smad2/3] (ab305325) at 1/1000 dilution

    Lane 1 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate 10 µg
    Lane 2 : C2C12 (mouse myoblasts myoblast), whole cell lysate 10 µg
    Lane 3 : C6 (rat glial tumor glial cell), whole cell lysate 10 µg

    Secondary
    All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

    Predicted band size: 48 kDa
    Observed band size: 58-62 kDa why is the actual band size different from the predicted?



    This data was developed using ab305325, the same antibody clone in a different buffer formulation. 

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 18809571).

    The identity of the band around 37 kDa is unknown.

    Exposure time: 59 seconds.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)

    This data was developed using ab305325, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Smad2 + Smad3 with ab305325 at 1/100 dilution (9.39 ug/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
    Positive staining on human colon is observed.
    The section was incubated with ab305325 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
    Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)

    This data was developed using ab305325, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling Smad2 + Smad3 with ab305325 at 1/100 dilution (9.39 ug/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
    Positive staining on human colon cancer is observed.
    The section was incubated with ab305325 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
    Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)

    This data was developed using ab305325, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Smad2 + Smad3 with ab305325 at 1/250 dilution (3.756 ug/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
    Positive staining on mouse colon is observed.
    The section was incubated with ab305325 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
    Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)

    This data was developed using ab305325, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded mouse breast cancer tissue labeling Smad2 + Smad3 with ab305325 at 1/250 dilution (3.756 ug/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
    Positive staining on mouse breast cancer is observed.
    The section was incubated with ab305325 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
    Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)
    Anti-Smad2 + Smad3 antibody [18/Smad2/3] - BSA and Azide free (ab305326)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (0)

Publishing research using ab305326? Please let us know so that we can cite the reference in this datasheet.

ab305326 has not yet been referenced specifically in any publications.

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