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    cd86-antibody-epr21962-ab239075.pdf

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Immunology Adaptive Immunity T Cells CD
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CD86 antibody [EPR21962] (ab239075)

  • Datasheet
  • SDS
  • Certificate of Compliance
Submit a review Submit a question References (4)

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Flow Cytometry - Anti-CD86 antibody [EPR21962] (ab239075)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)
  • Western blot - Anti-CD86 antibody [EPR21962] (ab239075)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)
  • Immunoprecipitation - Anti-CD86 antibody [EPR21962] (ab239075)
  • Western blot - Anti-CD86 antibody [EPR21962] (ab239075)
  • Anti-CD86 antibody [EPR21962] (ab239075)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21962] to CD86
  • Suitable for: WB, ICC/IF, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free

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Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Protein
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Recombinant human CD86 protein (Fc Chimera) (ab167720)
Primary
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Alexa Fluor® 488 Anti-IL-4 antibody [EPR1118Y] (ab211374)

View more associated products

Overview

  • Product name

    Anti-CD86 antibody [EPR21962]
    See all CD86 primary antibodies
  • Description

    Rabbit monoclonal [EPR21962] to CD86
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IPmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • Positive control

    • WB: Ramos, Raji and Daudi whole cell lysates. ICC/IF: Raji and Daudi cells. Flow Cyt: Raji cells.
  • General notes

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR21962
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Adaptive Immunity
    • T Cells
    • CD
    • Immunology
    • Cell Type Markers
    • CD
    • Non-lineage
    • Microbiology
    • Interspecies Interaction
    • Host Virus Interaction
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • B Lymphocytic Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Dendritic Cell Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Monocytic Lineage
    • Immunology
    • Adaptive Immunity
    • Regulatory T Cells
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • CD markers ELISA kits

Associated products

  • Alternative Versions

    • Anti-CD86 antibody [EPR21962] - BSA and Azide free (ab242020)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • ELISA pair antibody

    • Recombinant human CD86 protein (Fc Chimera) (ab167720)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human CD86 knockout Raji cell line (ab273858)
  • KO cell lysates

    • Human CD86 knockout Raji cell lysate (ab273812)
  • Recombinant Protein

    • Recombinant human CD86 protein (Fc Chimera) (ab167720)
  • Related Buffer

    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab239075 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
WB
1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 38 kDa).
ICC/IF
1/100.
Flow Cyt
1/500.
IP
1/30.
Notes
WB
1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 38 kDa).
ICC/IF
1/100.
Flow Cyt
1/500.
IP
1/30.
Application notes
Is unsuitable for IHC-P.

Target

  • Function

    Receptor involved in the costimulatory signal essential for T-lymphocyte proliferation and interleukin-2 production, by binding CD28 or CTLA-4. May play a critical role in the early events of T-cell activation and costimulation of naive T-cells, such as deciding between immunity and anergy that is made by T-cells within 24 hours after activation. Isoform 2 interferes with the formation of CD86 clusters, and thus acts as a negative regulator of T-cell activation.
  • Tissue specificity

    Expressed by activated B-lymphocytes and monocytes.
  • Sequence similarities

    Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Post-translational
    modifications

    Polyubiquitinated; which is promoted by MARCH8 and results in endocytosis and lysosomal degradation.
  • Cellular localization

    Membrane.
  • Target information above from: UniProt accession P42081 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 942 Human
    • Omim: 601020 Human
    • SwissProt: P42081 Human
    • Unigene: 171182 Human
    • Alternative names

      • Activation B7-2 antigen 3 antibody
      • Activation B7-2 antigen antibody
      • B-lymphocyte activation antigen B7-2 2 antibody
      • B-lymphocyte activation antigen B7-2 antibody
      • B7 2 antibody
      • B7 antibody
      • B7-2 antibody
      • B7.2 antibody
      • B70 antibody
      • B72 antibody
      • B72 antigen antibody
      • BU63 antibody
      • CD28 antigen ligand 2 2 antibody
      • CD28 antigen ligand 2 antibody
      • Cd28l2 antibody
      • CD28LG2 antibody
      • CD86 antibody
      • CD86 antigen (CD28 antigen ligand 2 B7 2 antigen) antibody
      • CD86 antigen (CD28 antigen ligand 2, B7-2 antigen) 1, 2 antibody
      • CD86 antigen (CD28 antigen ligand 2, B7-2 antigen) antibody
      • CD86 antigen antibody
      • CD86 molecule antibody
      • CD86_HUMAN antibody
      • CLS1 antibody
      • CTLA-4 counter-receptor B7.2 2 antibody
      • CTLA-4 counter-receptor B7.2 2, 3 antibody
      • CTLA-4 counter-receptor B7.2 antibody
      • Early T-cell costimulatory molecule 1 antibody
      • ETC-1 antibody
      • FUN 1 antibody
      • FUN-1 antibody
      • FUN1 antibody
      • LAB72 antibody
      • Ly-58 antibody
      • Ly58 antibody
      • MB7 antibody
      • MB7-2 antibody
      • Membrane glycoprotein antibody
      • MGC34413 antibody
      • T lymphocyte activation antigen CD86 precursor antibody
      • T-lymphocyte activation antigen CD86 antibody
      • TS/A-2 antibody
      see all

    Images

    • Flow Cytometry - Anti-CD86 antibody [EPR21962] (ab239075)
      Flow Cytometry - Anti-CD86 antibody [EPR21962] (ab239075)

      Flow cytometric analysis of Raji (human Burkitt's lymphoma cell line) cell line labeling CD86 with ab239075 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

      Gated on viable cells.

    • Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)
      Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Daudi (human Burkitt's lymphoma cell line) cells labeling CD86 with ab239075 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in Daudi cell line is observed.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Western blot - Anti-CD86 antibody [EPR21962] (ab239075)
      Western blot - Anti-CD86 antibody [EPR21962] (ab239075)
      All lanes : Anti-CD86 antibody [EPR21962] (ab239075) 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 - ab239075 observed at 70 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

      ab239075 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 ab239075 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.

    • Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)
      Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji (human Burkitt's lymphoma cell line) cells labeling CD86 with ab239075 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in Raji cell line is observed.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Immunoprecipitation - Anti-CD86 antibody [EPR21962] (ab239075)
      Immunoprecipitation - Anti-CD86 antibody [EPR21962] (ab239075)

      CD86 was immunoprecipitated from 0.35 mg of Raji whole (human Burkitt's lymphoma cell line) cell lysate with ab239075 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab239075 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

      Lane 1: Raji whole cell lysate 10 µg (Input). 

      Lane 2: ab239075 IP in Raji whole lysate. 

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab239075 in Raji whole cell lysate.

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

      Exposure time: 3 minutes.

    • Western blot - Anti-CD86 antibody [EPR21962] (ab239075)
      Western blot - Anti-CD86 antibody [EPR21962] (ab239075)
      All lanes : Anti-CD86 antibody [EPR21962] (ab239075) at 1/1000 dilution

      Lane 1 : Ramos (human Burkitt's lymphoma cell line) whole cell lysate
      Lane 2 : Raji (human Burkitt's lymphoma cell line) whole cell lysate
      Lane 3 : Daudi (human Burkitt's lymphoma cell line) whole cell lysate

      Lysates/proteins at 10 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?



      Exposure time : Lane 1: 3 minutes; Lanes 2-3: 26 seconds.

      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Anti-CD86 antibody [EPR21962] (ab239075)
      Anti-CD86 antibody [EPR21962] (ab239075)

    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

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (4)

    Publishing research using ab239075? Please let us know so that we can cite the reference in this datasheet.

    ab239075 has been referenced in 4 publications.

    • Chen Q  et al. EMP3 mediates glioblastoma-associated macrophage infiltration to drive T cell exclusion. J Exp Clin Cancer Res 40:160 (2021). PubMed: 33964937
    • Gu Y  et al. Enhancing Fatty Acid Catabolism of Macrophages Within Aberrant Breast Cancer Tumor Microenvironment Can Re-establish Antitumor Function. Front Cell Dev Biol 9:665869 (2021). PubMed: 33937269
    • Chen Y  et al. High Mobility Group Box 1 Contributes to the Acute Rejection of Liver Allografts by Activating Dendritic Cells. Front Immunol 12:679398 (2021). PubMed: 34177922
    • Zhang C  et al. miR-30c-5p Reduces Renal Ischemia-Reperfusion Involving Macrophage. Med Sci Monit 25:4362-4369 (2019). PubMed: 31185006

    Customer reviews and Q&As

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