B7-2/CD86 Antibody (BU63)

Western Blot: B7-2/CD86 Antibody (BU63) [NBP2-25208] - Western blot shows lysates of Ramos human Burkitt's lymphoma parental cell line and B7-2 knockout (KO) Ramos cell line. PVDF membrane was probed with 1.0 ug/mL of Mouse Anti-Human B7-2 Monoclonal Antibody (Catalog # NBP2-25208) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog #HAF018). Specific band was detected for B7-2 at approximately 90 kDa (as indicated) in the parental Ramos cell line, but is not detectable in the knockout Ramos cell line. This experiment was conducted under reducing conditions.

B7-2-CD86-Antibody-BU63-Immunohistochemistry-NBP2-25208-img0009.jpg

Immunocytochemistry/Immunofluorescence: B7-2/CD86 Antibody (BU63) [NBP2-25208] - Immunofluorescence staining of PFA-fixed Ramos cells using followed by goat anti-Mouse IgG conjugated to CF488 (green). Nuclei are stained with Red Dot.

B7-2-CD86-Antibody-BU63-Immunohistochemistry-NBP2-25208-img0010.jpg

Flow Cytometry: B7-2/CD86 Antibody (BU63) [NBP2-25208] - Flow Cytometric Analysis of PFA-fixed Ramos cells. B7-2/CD86 Antibody (BU63) followed by goat anti-Mouse IgG-CF488 (Blue); Isotype Control (Red).

Immunohistochemistry-Paraffin: B7-2/CD86 Antibody (BU63) [NBP2-25208] - Formalin-fixed, paraffin-embedded esophagus tumor tissue stained with CD86 antibody (5 ug/ml), peroxidase-conjugate and DAB chromogen. Note strong staining of differentiated squamous cells. TMA was used for this test.

Flow Cytometry: B7-2/CD86 Antibody (BU63) [NBP2-25208] - Flow Cytometric Analysis of human PBMCs using B7-2/CD86 Antibody (BU63); Goat anti-Mouse IgG-CF488 (red); Isotype Control (green).

Immunohistochemistry: B7-2/CD86 Antibody (BU63) [NBP2-25208] - Macrophage polarization states. Distribution & proportion of CD68+ (a), CD86+ (c) & CD163+ (e) macrophages (brown) were identified using IHC procedures. Intensity for the color response to antibodies was visible in lung tissues from all groups. Population proportions for the stained cells as shown by red arrows were calculated as a fold change of the control. (b, d, f). The results were expressed as Mean ± SD (n = 3). **: a p-value of < 0.01 vs NS, OVA/BUD & OVA/PCI except the number of CD86+ cells in OVA/PCI group. #: P < 0.05 vs NS & ##; P < 0.01 vs either NS or OVA/BUD group Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32111211), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: B7-2/CD86 Antibody (BU63) [NBP2-25208] - Effects of 20(S)-Rg3 on macrophage polarization in atherosclerotic lesions of diabetic ApoE–/– mice. (A,B) Co-immunofluorescence staining of the aortic root for macrophage (anti-Moma-2 antibody, green) & M1 marker iNos (red), & a bar graph summarizing the results (n = 5, respectively). (C,D) Co-immunofluorescence staining for macrophage (anti-Moma-2 antibody, green) & M1 marker CD86 (red), & a bar graph summarizing the results (n = 5, respectively). Scale bar: 20 μm. Data are mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29867472), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

M1 macrophages were induced in the in vivo mouse intervertebral disc (mIVD) degeneration model (puncture model). (a) Immunohistological analysis of Iba-1, iNOS, and arginase (brown staining) expression in nucleus pulposus (NP), annulus fibrosis (AF), and cartilage endplate (CEP) [right] at high magnification and in whole mIVDs (left) at low magnification. Images using an IgG isotype control antibody are included to demonstrate the specificity of the IHC staining. (b) Protein was extracted from punctured mIVDs in the puncture model, and western blotting was performed using antibodies against CD86, CD163, and beta -actin. (c) Images of the panel in (b), captured using a ChemiDoc Touch system and quantified using ImageJ. Biological and technical replicates were performed three times. Values represent the mean +/- SD. *p < 0.05, **p < 0.01, relative to the corresponding control. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/41053148), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Thrombin promotes polarization into M1 macrophages. (a) Mouse bone marrow was stimulated with thrombin, and quantitative PCR was performed using specific primers for CD86 and CD163. Biological and technical replicates were performed three times. (b) Mouse bone marrow was stimulated with thrombin, and western blotting was performed using antibodies for the M1 markers iNOS and CD86, and the M2 markers CD163 and Arginase-1. beta -actin was used as an internal control. n = 3. Values represent the mean +/- SD. *p < 0.05, **p < 0.01, relative to the corresponding control. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/41053148), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

IGU alleviates the inflammation of type II alveolar epithelial cells caused by M1 macrophages. (A–C) Representative western blot and quantitation for iNOS and CD86 in Raw264.7 cells. beta -actin was used as an internal control. (D) QRT-PCR for the mRNA levels of M1 macrophage polarization-related inflammatory cytokines TNF-alpha , IL-6 and IL-1 beta in Raw264.7 cells. (E, F) ELISA was used to measure the level of TNF-alpha and IL-6 in Raw264.7 cell culture supernatant. (G, H) QRT-PCR for the mRNA levels of MMP2 and MMP9. (I) Representative gelatin zymogram of MMPs in Raw264.7 cell culture supernatants. (J) Schematic representation of Raw264.7 cells co-cultured with type II alveolar epithelial cells. (K–M) QRT-PCR for the mRNA levels of TNF-alpha , IL-6 and IL-1 beta in type II alveolar epithelial cells (n=3). *P<0.05, **P<0.01, ***P<0.001, ns, No significance. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/40181990), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Rescue experiments of Raw264.7 cells after overexpression of TLR4. (A, B) Representative western blot and quantitation of TLR4 in TLR4-overexpressing Raw264.7 cells. beta -actin was used as an internal control. (C–E) QRT-PCR for the mRNA levels of M1 macrophage polarization-related inflammatory cytokines TNF-alpha , IL-6 and IL-1 beta in Raw264.7 cells. (F–J) Representative western blot and quantitation of protein levels of TLR4, NF-kappa B, iNOS and CD86 in Raw264.7 cells beta -actin was used as an internal control. (n=3). *P<0.05, **P<0.01, ***P<0.001, ns, No significance. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/40181990), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, Flow, ICC/IF, IHC, KO
• Clone: BU63
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 0.2 mg/ml

B7-2/CD86 Antibody (BU63) Summary

• Description: 200ug/ml of antibody purified from Bioreactor Concentrate by Protein A or G. Prepared in 10 mM PBS with 0.05% BSA & 0.05% azide. Also available WITHOUT BSA & azide at 1.0 mg/ml. (NBP2-34569)
  
  Antibody with azide - store at 2 to 8C. Antibody without azide - store at -20 to -80C.
• Additional Information: Clone BU63 was used by HLDA to establish CD designation.
• Immunogen: ARH-77 (B-lymphoblastoid cell line)
• Localization: Cell Surface
• Marker: Dendritic Cells Maturation Marker
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: CD86
• Purity: Protein A or G purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1-2 ug/million cells
  • Immunocytochemistry/ Immunofluorescence 1-2 ug/ml
  • Immunohistochemistry 0.5-1ug/ml
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 2-4 ug/ml
  • Knockout Validated
  • Simple Western 1:25
  • Western Blot 0.5-1ug/ml
• Application Notes: Immunohistochemistry (Formalin-fixed): 2-4ug/ml for 30 minutes at RT. Staining of formalin-fixed tissues requires heating tissue sections in 10mM Tris buffer with 1mM EDTA, pH 9.0, for 45 min at 95C followed by cooling at RT for 20 minutes.
  Optimal dilution for a specific application should be determined.
  IHC-fr reported in the literature (PMID: 29867472, 31629891)
  See Simple Western Antibody Database for Simple Western validation: antibody dilution of 1:25
• Theoretical MW: 70 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C.
• Buffer: 10 mM PBS with 0.05% BSA
• Preservative: 0.05% Sodium Azide
• Concentration: 0.2 mg/ml
• Purity: Protein A or G purified

Alternate Names for B7-2/CD86 Antibody (BU63)

• Activation B7-2 antigen
• B70
• B7-2 antigen
• B72
• B7-2
• B-lymphocyte activation antigen B7-2
• BU63
• CD28 antigen ligand 2
• CD28LG2B7-2 antigen)
• CD86 antigen
• CD86 molecule
• CD86
• CTLA-4 counter-receptor B7.2
• FUN-1
• LAB72
• MGC34413
• T-lymphocyte activation antigen CD86

Background

B7-2, also referred to as CD86, is a glycosylated type 1 membrane protein that is a member of the immunoglobulin (Ig) superfamily (1). B7-2/CD86 is constitutively expressed on antigen presenting cells (APCs) such as dendritic cells (DCs), macrophages, and B cells and functions in controlling immune responses through either costimulatory or coinhibitory signals (1,2). Expression of B7-2 is upregulated by APCs upon activation and can be induced in T cells (1,3). The human B7-2 protein, encoded by the CD86 gene, is 329 amino acids (aa) in length with a theoretical molecular weight (MW) of 37.6 kDa (4). The B7-2 protein contains characteristic Ig variable-like (IgV) and constant-like (IgC) domains within its extracellular region (1,3). B7-2/CD86 has structural similarity and shares ~25% sequence homology with another B7 family molecule, B7-1/CD80 (3,5). Both B7-1 and B7-2 are ligands for the activating receptor CD28 and the regulatory receptor cytotoxic T-lymphocyte antigen 4 (CTLA-4) which are expressed on subsets of T cells (1-3,5-7). However, B7-1 and B7-2 bind to CTLA-4 with higher affinity than CD28 (3,5,6). B7-2/CD86 binding to CD28 results in costimulatory signals to promote T cell activation, proliferation, and cytokine production (1-3. 5-7). Binding to CTLA-4 initiates coinhibitory signaling to attenuate the pro-inflammatory T cell response, while also promoting the suppressive function of regulatory T (Treg) cells through expression of indoleamine 2,3-deoxygenase (IDO) (1-3,6,7). Molecules involved in T cell co-signaling have become considerable targets of interest for cancer immunotherapy (7). The anti-CTLA-4 monoclonal antibody ipilimumab, first approved for the treatment of metastatic melanoma, prevents B7-1/B7-2 molecules from binding to CTLA-4, thus driving B7-CD28 binding and promoting costimulatory signals and antitumor effects (1,7,8). Combination therapies targeting co-signaling molecules are currently under investigation to improve antitumor response for treatment of both melanoma and non-melanoma cancers (7,8).

References

1. Collins M, Ling V, Carreno BM. The B7 family of immune-regulatory ligands. Genome Biol. 2005;6(6):223. https://doi.org/10.1186/gb-2005-6-6-223

2. Greaves P, Gribben JG. The role of B7 family molecules in hematologic malignancy. Blood. 2013;121(5):734-744. https://doi.org/10.1182/blood-2012-10-385591

3. Bolandi N, Derakhshani A, Hemmat N, et al. The Positive and Negative Immunoregulatory Role of B7 Family: Promising Novel Targets in Gastric Cancer Treatment. Int J Mol Sci. 2021;22(19):10719. https://doi.org/10.3390/ijms221910719

4. Uniprot (P42081)

5. Bhatia S, Edidin M, Almo SC, Nathenson SG. B7-1 and B7-2: similar costimulatory ligands with different biochemical, oligomeric and signaling properties. Immunol Lett. 2006;104(1-2):70-75. https://doi.org/10.1016/j.imlet.2005.11.019

6. Ohue Y, Nishikawa H. Regulatory T (Treg) cells in cancer: Can Treg cells be a new therapeutic target?. Cancer Sci. 2019;110(7):2080-2089. https://doi.org/10.1111/cas.14069

7. Chen L, Flies DB. Molecular mechanisms of T cell co-stimulation and co-inhibition [published correction appears in Nat Rev Immunol. 2013 Jul;13(7):542]. Nat Rev Immunol. 2013;13(4):227-242. https://doi.org/1010.1038/nri3405

8. Karimi A, Alilou S, Mirzaei HR. Adverse Events Following Administration of Anti-CTLA4 Antibody Ipilimumab. Front Oncol. 2021;11:624780. https://doi.org/101010.3389/fonc.2021.624780

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for B7-2/CD86 Antibody (NBP2-25208)(34)

Publications using NBP2-25208

• Wang B, Chen Y, Chang H et al. Zinc Oxide Nanoparticles Exacerbate Skin Epithelial Cell Damage by Activating M1 Macrophages by Upregulating the Nlrp3 Inflammasome and Exosome Secretion after Uvb Irradiation-Induced Skin Injury SSRN Electronic Journal 2023-02-22 (IHC-P, Mouse)
• W B, Chen Y, Chang H et al. Zinc oxide nanoparticles exacerbate skin epithelial cell damage by upregulating the NLRP3 inflammasome and exosome secretion in M1 macrophages after UVB irradiation-induced skin injury Research Square 2023-09-15 (IHC-P, Mouse)
• Zhao Q, Zhang Y, Wen K et al. Myeloid but not hepatocytic CD38 is a key driver for hepatic ischemia/reperfusion injury. Signal transduction and targeted therapy 2025-05-09 [PMID: 40341132]
• Li K, Xu W, Chen Y et al. Piezoelectric Nanostructured Surface for Ultrasound?Driven Immunoregulation to Rescue Titanium Implant Infection Advanced Functional Materials 2023-04-18
• Tatsuno R, Ando T, Fujimaki T et al. Thrombin induces degradation of murine intervertebral discs via angiogenesis and M1-like polarization of macrophages Scientific reports 2025-10-06 [PMID: 41053148]
• Lu Z, Chai Q, Dai W et al. Mitochondrial homeostasis restoring peptide-drug conjugates with ROS-responsive NO releasing ability for targeted therapy of myocardial infarction. Journal of Nanobiotechnology 2025-07-09 [PMID: 40629337]
• Zhu W, Chang J, Tian L et al. Depletion of HSP60 in Microglia Leads to Synaptic Dysfunction and Depression-Like Behaviors Through Enhanced Synaptic Pruning in Male Mice. CNS neuroscience & therapeutics 2025-04-16 [PMID: 40237297]
• Xia Z, Zhou C, Hong Y et Al. TFPI2 hypermethylation promotes diabetic atherosclerosis progression through the Ap2?/PPAR? axis J Mol Cell Cardiol 2024-12-03 [PMID: 39631358]
• Liu J, Shi Y, Zhao Y et Al. A Multifunctional Metal-Phenolic Nanocoating on Bone Implants for Enhanced Osseointegration via Early Immunomodulation Adv Sci (Weinh) 2024-03-06 [PMID: 38445899]
• Prinz LF, Riet T, Neureuther DF, Lennartz S et Al. An anti-CD19/CTLA-4 switch improves efficacy and selectivity of CAR T cells targeting CD80/86-upregulated DLBCL Cell Rep Med 2024-02-10 [PMID: 38340727]
• Shi C, Zhang Y, Wu G et al. Hyaluronic Acid-Based Reactive Oxygen Species-Responsive Multifunctional Injectable Hydrogel Platform Accelerating Diabetic Wound Healing Advanced healthcare materials 2023-11-09 [PMID: 37943252]
• Luo Y, Ali T, Liu Z et al. EPO prevents neuroinflammation and relieves depression via JAK/STAT signaling Life Sciences 2023-09-01 [PMID: 37769806] (WB, Mouse)
• Jia H, Wei P, Zhou S et al. Attenuated Salmonella carrying siRNA-PD-L1 and radiation combinatorial therapy induces tumor regression on HCC through T cell-mediated immuno-enhancement Cell Death Discovery 2023-08-28 [PMID: 37640735] (Western Blot, Human)
• Tan Y, Coyle RC, Barrs RW et al. Nanowired human cardiac organoid transplantation enables highly efficient and effective recovery of infarcted hearts Science Advances 2023-08-04 [PMID: 37540743] (Western Blot, Human)
• Chen S, Xing Z, Geng M et al. Macrophage fusion event as one prerequisite for inorganic nanoparticle-induced antitumor response Science advances 2023-07-21 [PMID: 37467339] (IHC-P, Mouse)
• Yao S, Zhou Z, Wang L et al. Targeting endometrial inflammation in intrauterine adhesion ameliorates endometrial fibrosis by priming MSCs to secrete C1INH iScience 2023-06-01 [PMID: 37456855]
• Chen L, Song X, Yao Z et al. Gelatin nanofiber-reinforced decellularized amniotic membrane promotes axon regeneration and functional recovery in the surgical treatment of peripheral nerve injury Biomaterials 2023-09-01 [PMID: 37352606] (IHC-P)
• Tan X, Fang P, Li K et al. A HER2-targeted antibody-novel DNA topoisomerase I inhibitor conjugate induces durable adaptive antitumor immunity by activating dendritic cells mAbs 2023-06-14 [PMID: 37314961] (IHC-P, Human)
  
  Details:
  1:200 dilution
• Ling Q, Fang J, Zhai C et al. Berberine induces SOCS1 pathway to reprogram the M1 polarization of macrophages via miR-155-5p in colitis-associated colorectal cancer European journal of pharmacology 2023-04-13 [PMID: 37059377]
• Jia X, Xie L, Liu y et al. Astragalus polysaccharide (APS) exerts protective effect against acute ischemic stroke (AIS) through enhancing M2 micoglia polarization by regulating adenosine triphosphate (ATP)/ purinergic receptor (P2X7R) axis Bioengineered 2022-02-01 [PMID: 35166175] (IHC-P, Rat)
• Luo ZW, Xia K, Liu YW et al. Extracellular Vesicles from Akkermansia muciniphila Elicit Antitumor Immunity Against Prostate Cancer via Modulation of CD8+ T Cells and Macrophages International journal of nanomedicine 2021-04-20 [PMID: 33907401] (IF/IHC, Human)
• Liu P, Peng C, Chen X et al. Acitretin Promotes the Differentiation of Myeloid-Derived Suppressor Cells in the Treatment of Psoriasis Frontiers in Medicine 2021-03-23 [PMID: 33834031] (IHC-P, Mouse)
• Yang D, Shen L, Chen R et al. The Effect of Talaromyces Marneffei Infection on CD86 Expression in THP-1 Cells Infect Drug Resist 2021-03-01 [PMID: 33642869]
• Klinge U, Dievernich A, Tolba R et al. CD68+ macrophages as crucial components of the foreign body reaction demonstrate an unconventional pattern of functional markers quantified by analysis with double fluorescence staining J. Biomed. Mater. Res. Part B Appl. Biomater. 2020-05-30 [PMID: 32475069] (ICC/IF, Human)
• Li Ml, Su Xm, Ren Y et Al. HDAC8 inhibitor attenuates airway responses to antigen stimulus through synchronously suppressing galectin-3 expression and reducing macrophage-2 polarization Respir. Res. 2020-02-28 [PMID: 32111211] (IF/IHC, Mouse)
• Shehata MM, Saad Eldien HM, Meligy FY, Bahaidarh SY The Possible Protective Role of Barley Seeds on the Spleen after Administration of Glucocorticoids in Adult Albino Rats: A Histological and Immunohistochemical Study J Microsc Ultrastruct 2019-11-18 [PMID: 31803571] (Rat)
• Joly-Amado A, Davtyan H, Serraneau K et al. Active immunization with tau epitope in a mouse model of tauopathy induced strong antibody response together with improvement in short memory and pSer396-tau pathology. Neurobiol Dis. 2020-02-01 [PMID: 31629891] (IHC-Fr, Mouse)
• Zhu LF, Li L, Wang XQ et al. M1 macrophages regulate TLR4/AP1 via paracrine to promote alveolar bone destruction in periodontitis Oral Dis 2019-07-30 [PMID: 31361069]
• Liu D, Guo M, Zhou P et al. TSLP promote M2 macrophages polarization and cardiac healing after myocardial infarction Biochem. Biophys. Res. Commun. 2019-06-18 [PMID: 31227217] (IF, Mouse)
• Kaushik NK, Kaushik N, Adhikari M et al. Preventing the Solid Cancer Progression via Release of Anticancer-Cytokines in Co-Culture with Cold Plasma-Stimulated Macrophages Cancers (Basel) 2019-06-18 [PMID: 31216715] (IHC-P, Mouse)
• Zhao J, Zhang Z, Xue Y et al. Anti-tumor macrophages activated by ferumoxytol combined or surface-functionalized with the TLR3 agonist poly (I : C) promote melanoma regression Theranostics 2018-11-29 [PMID: 30613299] (ICC/IF, Human)
• Rivera Del Alamo M, Diaz-Lobo M, Busquets S et al. Specific expression pattern of tissue cytokines analyzed through the Surface Acoustic Wave technique is associated with age-related spontaneous benign prostatic hyperplasia in rats. Biochem Biophys Rep 2018-01-01 [PMID: 29872731] (WB, Rat)
• Ginsenoside Rg3 Mitigates Atherosclerosis Progression in Diabetic apoE-/- Mice by Skewing Macrophages to the M2 Phenotype. Ginsenoside Rg3 Mitigates Atherosclerosis Progression in Diabetic apoE-/- Mice by Skewing Macrophages to the M2 Phenotype. Front Pharmacol. 2018-05-09 [PMID: 29867472] (IHC-Fr, Mouse)
• Schliefsteiner C, Peinhaupt M, Kopp S et al. Human Placental Hofbauer Cells Maintain an Anti-inflammatory M2 Phenotype despite the Presence of Gestational Diabetes Mellitus Front Immunol 2017-08-21 [PMID: 28824621] (WB, Human)

Review for B7-2/CD86 Antibody (NBP2-25208) (1)

Average Rating: 3

(Based on 1 review)

We have 1 review tested in 1 species: Rat.

reviewed by: Sophie E IHC-P Rat 07/03/2025

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: Rat Lung Tissue
• Species: Rat
• Lot: 942-1P220621

Comments

• Comments: Native lung tissue (Rat) stained with CD86 (green) and Elastin (Eln) (red). Fixed in 4% PFA then paraffin-embedded and sectioned.

Bioinformatics

• Gene Symbol: CD86
• Uniprot:
  • P42081()
  • P42081()
  • NP_001193853(Human)