Recombinant
RabMAb

Anti-CD86 antibody [EP1158Y] (ab53004)

Overview

  • Product name
    Anti-CD86 antibody [EP1158Y]
    See all CD86 primary antibodies
  • Description
    Rabbit monoclonal [EP1158Y] to CD86
  • Specificity
    This antibody has been found to work on mouse samples in WB but not IHC.
  • Tested applications
    Suitable for: WB, IP, ICC, Flow Cyt, IHC-P, ICC/IF, IHC-Frmore details
  • Species reactivity
    Reacts with: Human, Cynomolgus monkey
  • Immunogen

    synthetic peptide corresponding to residues near the N-terminus of human B7-2 was used as an immunogen

  • Positive control
    • Raji cell membrane lysate Human tonsil tissue
  • 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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab53004 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 1/5000. Detects a band of approximately 70 kDa.
IP 1/50.
ICC 1/100 - 1/250.
Flow Cyt 1/10.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P Use at an assay dependent concentration.
ICC/IF 1/500.
IHC-Fr Use at an assay dependent concentration. PubMed: 21998115

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.
  • Information by UniProt
  • Database links
  • 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 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) 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, 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
    • LAB72 antibody
    • Ly-58 antibody
    • Ly58 antibody
    • MB7 antibody
    • MB7-2 antibody
    • Membrane glycoprotein antibody
    • MGC34413 antibody
    • T-lymphocyte activation antigen CD86 antibody
    • TS/A-2 antibody
    see all

Images

  • ab53004 staining CD86 in Cynomolgus Monkey lymph node tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 10% serum for 20 minutes at room temperature; antigen retrieval was by heat mediation in a citrate buffer, pH 6.0. Samples were incubated with primary antibody (1/800) for 30 minutes at room temperature. An undiluted Biotin-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

  • Immunocytochemistry/Immunofluorescence analysis of Raji (Human Burkitt's lymphoma cell line) labelling CD86 with purified ab53004 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% triton X-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

  • Anti-CD86 antibody [EP1158Y] (ab53004) at 1/5000 dilution + Raji cell lysate at 10 µg

    Secondary
    goat anti-rabbit HRP labelled at 1/2000 dilution

    Observed band size : 70 kDa (why is the actual band size different from the predicted?)
  • ab53004, at a 1/100 dilution, staining Human CD86 in Tonsil, using Immunohistochemistry, Paraffin embedded tissue.
  • Overlay histogram showing Raji cells stained with ab53004 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab53004, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive result in 80% methanol (5 min) fixed Raji cells used under the same conditions.

    Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

References

This product has been referenced in:
  • Subramanian S  et al. TREM-1 associated macrophage polarization plays a significant role in inducing insulin resistance in obese population. J Transl Med 15:85 (2017). IHC-P ; Human . Read more (PubMed: 28454543) »
  • Deng Y  et al. Injectable in situ cross-linking chitosan-hyaluronic acid based hydrogels for abdominal tissue regeneration. Sci Rep 7:2699 (2017). IF . Read more (PubMed: 28578386) »

See all 16 Publications for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Horse Tissue sections (Lymph node)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization
No
Specification
Lymph node
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jun 24 2016

Application
Immunohistochemistry (Frozen sections)
Sample
Dog Tissue sections (spleen)
Permeabilization
No
Specification
spleen
Blocking step
Serum as blocking agent for 1 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Acetone
Username

Abcam user community

Verified customer

Submitted Feb 06 2016

Application
Immunohistochemistry (Frozen sections)
Sample
Horse Tissue sections (spleen)
Permeabilization
No
Specification
spleen
Blocking step
Serum as blocking agent for 1 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Acetone
Username

Abcam user community

Verified customer

Submitted Feb 06 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA pH8.0
Permeabilization
No
Specification
Lung
Blocking step
(agent) for 4 minute(s) · Concentration: 100% · Temperature: 36°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jun 23 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Mammary tumor (MMATV-PyMT))
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 5 min high pressure
Permeabilization
Yes - 1mM EDTA, 0.05% Tween 20, pH 8.0
Specification
Mammary tumor (MMATV-PyMT)
Blocking step
5% goat serum +3% BSA in IF buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Apr 14 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
1% BSA + 5% NGS + 0.1% Sapo as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 20°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Sample
Human Tissue sections (blood vessels)
Specification
blood vessels
Permeabilization
Yes - saponine
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 24 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Sample
Rat Tissue sections (spleen)
Specification
spleen
Permeabilization
No
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Dec 04 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: Rt°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric buffer pH 6.0
Sample
Cynomolgus Monkey Tissue sections (Lymph node)
Specification
Lymph node
Permeabilization
No
Fixative
Formaldehyde
Username

Jing Ma

Verified customer

Submitted Sep 02 2013

Thank you for contacting us.

The links to the entry for CD86 in the Human ProteinAtlas are as follows:

http://www.proteinatlas.org/ENSG00000114013
http://www.proteinatlas.org/ENSG00000114013/normal (specifically for the normal tis...

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I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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