Recombinant 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
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 within Human CD86 aa 1-250. The exact sequence is proprietary.
Database link: P42081 -
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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, as well as 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... -
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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Recombinant Protein
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Related Buffer
Applications
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. |
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)](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)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
Certificate of Compliance
References (1)
ab239075 has been referenced in 1 publication.
- Zhang C et al. miR-30c-5p Reduces Renal Ischemia-Reperfusion Involving Macrophage. Med Sci Monit 25:4362-4369 (2019). PubMed: 31185006
