Recombinant Anti-CD86 antibody [EPR21962] (ab239075)
![Flow Cytometry - Anti-CD86 antibody [EPR21962] (ab239075)](https://www.abcam.com/ps/products/239/ab239075/Images/ab239075-326984-anti-cd86-antibody-epr21962-flow-cytometry.jpg "Flow Cytometry - 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
Related conjugates and formulations
Overview
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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.
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Positive control
- WB: Ramos, Raji and Daudi whole cell lysates. ICC/IF: Raji and Daudi cells. Flow Cyt: Raji cells.
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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
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
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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... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21962 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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ELISA pair antibody
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Buffer
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).
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| ICC/IF |
1/100.
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| Flow Cyt |
1/500.
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| IP |
1/30.
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| Notes |
|---|
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WB
1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 38 kDa). |
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ICC/IF
1/100. |
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Flow Cyt
1/500. |
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IP
1/30. |
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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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.
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![Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)](https://www.abcam.com/ps/products/239/ab239075/Images/ab239075-326987-anti-cd86-antibody-epr21962-immunofluorescence.jpg)
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.
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![Western blot - Anti-CD86 antibody [EPR21962] (ab239075)](https://www.abcam.com/ps/products/239/ab239075/Images/ab239075-448036-anti-cd86-antibody-epr21962-western-blot-raji-wt-raji-cd86-knockout.jpg)
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.
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![Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)](https://www.abcam.com/ps/products/239/ab239075/Images/ab239075-326986-anti-cd86-antibody-epr21962-immunofluorescence.jpg)
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.
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![Immunoprecipitation - Anti-CD86 antibody [EPR21962] (ab239075)](https://www.abcam.com/ps/products/239/ab239075/Images/ab239075-326985-anti-cd86-antibody-epr21962-immunoprecipitation.jpg)
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.
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![Western blot - Anti-CD86 antibody [EPR21962] (ab239075)](https://www.abcam.com/ps/products/239/ab239075/Images/ab239075-326983-anti-cd86-antibody-epr21962-western-blot.jpg)
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.
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![Anti-CD86 antibody [EPR21962] (ab239075)](https://www.abcam.com/ps/products/239/ab239075/Images/ab239075-6-benefits-of-recombinant-antibodies.png)
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.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (4)
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
