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
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Product name
Anti-CD86 antibody [EP1158-37]
See all CD86 primary antibodies -
Description
Rabbit monoclonal [EP1158-37] to CD86 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Raji, Daudi, Jurkat, Human tonsil and Human spleen lysates. IHC-P: Human non-Hodgkin lymphoma and Human tonsil tissues.
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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.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1158-37 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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KO cell lines
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KO cell lysates
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab269587 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 |
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WB |
1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 38 kDa).
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IHC-P |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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WB
1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 38 kDa). |
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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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
modificationsPolyubiquitinated; which is promoted by MARCH8 and results in endocytosis and lysosomal degradation. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 942 Human
- Omim: 601020 Human
- SwissProt: P42081 Human
- Unigene: 171182 Human
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Alternative names
- Activation B7-2 antigen 3 antibody
- Activation B7-2 antigen antibody
- B-lymphocyte activation antigen B7-2 2 antibody
see all
Images
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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.
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All lanes : Anti-CD86 antibody [EP1158-37] (ab269587) at 1/1000 dilution
Lane 1 : Wild-type Raji (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 2 : CD86 knockout Raji (Human Burkitt's lymphoma cell line) whole 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.
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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).
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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.
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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.
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (5)
ab269587 has been referenced in 5 publications.
- Zhou Y et al. Upregulation of MTA1 in Colon Cancer Drives A CD8+ T Cell-Rich But Classical Macrophage-Lacking Immunosuppressive Tumor Microenvironment. Front Oncol 12:825783 (2022). PubMed: 35350571
- Zhang M et al. The Distributional Characteristics of M2 Macrophages in the Placental Chorionic Villi are Altered Among the Term Pregnant Women With Uncontrolled Type 2 Diabetes Mellitus. Front Immunol 13:837391 (2022). PubMed: 35386706
- Yin XK et al. Expression Pattern and Prognostic Value of CTLA-4, CD86, and Tumor-Infiltrating Lymphocytes in Rectal Cancer after Neoadjuvant Chemo(radio)therapy. Cancers (Basel) 14:N/A (2022). PubMed: 36428666
- Macedo C et al. Donor-derived regulatory dendritic cell infusion results in host cell cross-dressing and T cell subset changes in prospective living donor liver transplant recipients. Am J Transplant 21:2372-2386 (2021). PubMed: 33171019
- Borcherding L et al. Impaired Dendritic Cell Homing in COVID-19. Front Med (Lausanne) 8:761372 (2021). PubMed: 34805226