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Anti-ENPP2/ATX antibody [1F8] - BSA and Azide free (ab238670)

Key features and details

  • Mouse monoclonal [1F8] to ENPP2/ATX - BSA and Azide free
  • Suitable for: WB, ELISA, IHC-P, ICC/IF
  • Reacts with: Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-ENPP2/ATX antibody [1F8] - BSA and Azide free
    See all ENPP2/ATX primary antibodies

  • Description

    Mouse monoclonal [1F8] to ENPP2/ATX - BSA and Azide free

  • Host species

    Mouse

  • Tested applications

    Suitable for: WB, ELISA, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse

  • Immunogen

    Recombinant full length protein corresponding to Human ENPP2/ATX.

  • Positive control

    • WB: HEK293 whole cell lysate and in the following human tissue lysates: kidney; placenta; ovary; small intestine. IHC: human tonsil paraffin sections. ICC/IF: HeLa cells.

  • General notes

    ab238670 is a PBS only version of ab77104.

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Images

  • Western blot - Anti-ENPP2 antibody [5H3] - BSA and Azide free (ab238670)
    Western blot - Anti-ENPP2 antibody [5H3] - BSA and Azide free (ab238670)
    All lanes : Anti-ENPP2/ATX antibody [1F8] (ab77104) at 10 µg/ml

    Lane 1 : ab30203
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 99 kDa
    Observed band size: 100 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 180 kDa, 250 kDa, 50 kDa, 65 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes


    This data was developed using the same antibody clone in a different formulation containing PBS, Azide and arginine (ab77104).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENPP2 antibody [5H3] - BSA and Azide free (ab238670)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENPP2 antibody [5H3] - BSA and Azide free (ab238670)

    IHC image of ENPP2/ATX staining in Human Tonsil FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab77104, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    This data was developed using the same antibody clone in a different formulation containing PBS, Azide and arginine (ab77104).

  • Immunocytochemistry/ Immunofluorescence - Anti-ENPP2 antibody [5H3] - BSA and Azide free (ab238670)
    Immunocytochemistry/ Immunofluorescence - Anti-ENPP2 antibody [5H3] - BSA and Azide free (ab238670)

    ICC/IF image of ab77104 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab77104, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    This data was developed using the same antibody clone in a different formulation containing PBS, Azide and arginine (ab77104).

  • Western blot - Anti-ENPP2 antibody [5H3] - BSA and Azide free (ab238670)
    Western blot - Anti-ENPP2 antibody [5H3] - BSA and Azide free (ab238670)
    All lanes : Anti-ENPP2/ATX antibody [1F8] (ab77104) at 10 µg/ml

    Lane 1 : ab29745
    Lane 2 : ab30222
    Lane 3 : ab29276

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Predicted band size: 99 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?
    Additional bands at: 300 kDa, 40 kDa, 55 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes


    ENPP2/ATX contains three potential glycosylation sites (SwissProt), which might explain its migration at a higher molecular weight than predicted.

    This data was developed using the same antibody clone in a different formulation containing PBS, Azide and arginine (ab77104).

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