Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CD86 antibody [EP1158-37] (ab269587)

Key features and details

  • Rabbit monoclonal [EP1158-37] to CD86
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG

Overview

  • Product name

    Anti-CD86 antibody [EP1158-37]
    See all CD86 primary antibodies

  • Description

    Rabbit monoclonal [EP1158-37] to CD86

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: Raji, Daudi, Jurkat, Human tonsil and Human spleen lysates. IHC-P: Human non-Hodgkin lymphoma and Human tonsil tissues.

  • 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.

Properties

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD86 antibody [EP1158-37] (ab269587)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD86 antibody [EP1158-37] (ab269587)

    Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling CD86 with ab269587 at 1/100 dilution (2.52 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human tonsil (PMID: 22740445). The section was incubated with ab269587 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

  • Western blot - Anti-CD86 antibody [EP1158-37] (ab269587)
    Western blot - Anti-CD86 antibody [EP1158-37] (ab269587)
    All lanes : Anti-CD86 antibody [EP1158-37] (ab269587) at 1/1000 dilution

    Lane 1 : Wild-type Raji cell lysate
    Lane 2 : CD86 knockout Raji cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 38 kDa
    Observed band size: 70 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 2: Merged signal (red and green). Green - ab269587 observed at 70 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

    ab269587 was shown to react with CD86 in wild-type Raji cells in western blot with loss of signal observed in CD86 knockout sample. Wild-type and CD86 knockout Raji cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab269587 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] 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.

  • Western blot - Anti-CD86 antibody [EP1158-37] (ab269587)
    Western blot - Anti-CD86 antibody [EP1158-37] (ab269587)
    All lanes : Anti-CD86 antibody [EP1158-37] (ab269587) at 1/1000 dilution

    Lane 1 : Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate
    Lane 2 : Daudi (human burkitt's lymphoma lymphoblast) whole cell lysate
    Lane 3 : Jurkat (human t cell leukemia t lymphocyte) 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

    Predicted band size: 38 kDa
    Observed band size: 80 kDa why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBSt.

    Exposure time: 3 minutes.

    Negative control: Jurkat (PMID: 9973476).

    The actual band size higher than predicted is mainly due to glycosylation.

    The molecular weight observed is consistent with what has been described in the literature (PMID:27464357, 21896490).

  • Western blot - Anti-CD86 antibody [EP1158-37] (ab269587)
    Western blot - Anti-CD86 antibody [EP1158-37] (ab269587)
    All lanes : Anti-CD86 antibody [EP1158-37] (ab269587) at 1/1000 dilution

    Lane 1 : Human tonsil lysate
    Lane 2 : Human spleen lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

    Predicted band size: 38 kDa
    Observed band size: 80 kDa why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    The actual band size higher than predicted is mainly due to glycosylation.

    The molecular weight observed is consistent with what has been described in the literature (PMID:27464357, 21896490).

    This blot was developed using a higher sensitivity ECL substrate.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD86 antibody [EP1158-37] (ab269587)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD86 antibody [EP1158-37] (ab269587)

    Immunohistochemical analysis of paraffin-embedded Human non-Hodgkin lymphoma tissue labeling CD86 with ab269587 at 1/100 dilution (2.52 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human non-Hodgkin lymphoma (PMID: 9767448,10089910). The section was incubated with ab269587 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

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