Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rat monoclonal [CD3-12] to CD3 - BSA and Azide free
  • Suitable for: IHC-P, WB, Flow Cyt
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-CD3 antibody [CD3-12] - BSA and Azide free
    See all CD3 primary antibodies

  • Description

    Rat monoclonal [CD3-12] to CD3 - BSA and Azide free

  • Host species

    Rat

  • Tested applications

    Suitable for: IHC-P, WB, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • IHC-P: Human tonsil tissue. Mouse spleen and lymph node tissue; Rat spleen tissue. Flow Cyt: Human peripheral blood lymphocytes. WB: Jurkat, MOLT-4 and EL4 cell lysates; Human, mouse and rat thymus tissue lysates.

  • General notes

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

    ab255972 is the carrier-free version of ab11089. 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

    This product 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.

Properties

Images

  • Flow Cytometry - Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)
    Flow Cytometry - Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab11089).

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 0.1% Tween-20 permeabilized human peripheral blood mononuclear cells (PBMC) labeling CD3 with ab11089 at 1/400 dilution (red) compared with a Mouse IgG, monoclonal Isotype Control (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Mouse IgG ab150157 (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • Western blot - Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)
    Western blot - Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)
    All lanes : Anti-CD3 antibody [CD3-12] (ab11089) at 1 µg/ml

    Lane 1 : THP-1 (Human monocytic leukemia cell line) whole cell lysate (negative control)
    Lane 2 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate (negative control)
    Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 4 : Human thymus tissue lysate
    Lane 5 : Mouse thymus tissue lysate
    Lane 6 : Rat thymus tissue lysate

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat anti-Rat at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 19 kDa
    Observed band size: 23 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 containing PBS, BSA, glycerol, and sodium azide (ab11089).

    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab11089 overnight at 4°C. Antibody binding was detected using Goat anti-Rat secondary at a 1:10000 dilution for 1hr at room temperature and then imaged.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab11089).

    Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CB3 with ab11089 at 1/200 dilution, followed by ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882). Positive staining on rat spleen tissue is observed. Counterstained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use Goat Anti-Mouse IgG H&L (HRP polymer) (ab214882). Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab11089).

    Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD3 with ab11089 at 1/200 dilution, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Positive staining on mouse spleen tissue is observed. Counterstained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use Goat Anti-Mouse IgG H&L (HRP polymer) (ab214882). Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab11089).

    IHC image of CD3 staining in a formalin fixed, paraffin embedded human tonsil tissue. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH 6). The section was incubated with ab11089 at 1/250 dilution for 15 minutes at room temperature. A goat anti-rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. The section was counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab11089).

    IHC image of CD3 staining in a formalin fixed, paraffin embedded normal mouse lymph node tissue. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH 6). The section was incubated with ab11089 at 1/250 dilution for 15 minutes at room temperature. A goat anti-rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. The section was counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Western blot - Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)
    Western blot - Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)
    All lanes : Anti-CD3 antibody [CD3-12] (ab11089) at 1/1000 dilution

    Lane 1 : Mouse thymus tissue lysate
    Lane 2 : Rat thymus tissue lysate
    Lane 3 : Human thymus tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

    Predicted band size: 19 kDa



    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab11089).

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure: Lanes 1/3: 5 secs; Lanes 2: 3 secs.

  • Western blot - Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)
    Western blot - Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)
    All lanes : Anti-CD3 antibody [CD3-12] (ab11089) at 1/1000 dilution

    Lane 1 : Jurkat (human T cell leukemia T lymphocyte)
    Lane 2 : MOLT-4 (human lymphoblastic leukemia T lymphoblast)
    Lane 3 : EL4 (mouse lymphoma T lymphocyte), whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

    Predicted band size: 19 kDa



    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab11089).

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 5 secs.

  • Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)
    Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)

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