PA007620.r2b: In Vivo Grade Recombinant Anti-mouse CD86 (B7-2) Monoclonal Antibody (Clone: 2D10), Rat IgG2b Kappa
Recombinant Rat IgG2b Monoclonal Antibody.
Clone: 2D10.
Isotype: Rat IgG2b Kappa.
Source: The anti-mouse CD86 (B7-2) monoclonal antibody (clone: 2D10) was produced in mammalian cells.
Specificity/Sensitivity: The in vivo grade recombinant rat monoclonal antibody (clone: 2D10) specifically binds to mouse CD86 (B7-2).
Applications: ELISA, neutralization, functional assays such as bioanalytical PK and ADA assays, and those assays for studying biological pathways affected by the mouse CD86 (B7-2) protein.
Form of Antibody: 0.2 uM filtered solution, pH 7.4, no stabilizers or preservatives.
Endotoxin: < 1 EU per 1 mg of the protein by the LAL method.
Purity: >95% by SDS-PAGE under reducing conditions and HPLC.
Shipping: The in vivo grade recombinant anti-mouse CD86 (B7-2) monoclonal antibody of clone 2D10 is shipped with ice pack. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70°C as supplied.
1 month from date of receipt, 2 to 8°C as supplied.
References of anti-mouse CD86 (B7-2) antibody (Clone: 2D10):
Toll-like Receptor 2 Facilitates Oxidative Damage-Induced Retinal Degeneration
Radu, C. G., et al. Invest Ophthalmol Vis Sci. 2020 May 11;61(5):2. doi: 10.1167/iovs.61.5.2. PMID: 32350575
PBMCs were labeled for the investigation of monocytes with the following fluorochrome-labeled antibodies: anti-CD16 (3G8) anti-CD86 (FM95) anti-CD45 (2D1); anti-CD66b (G10F5); CD14 (Tük4); CD80 (2D10); (Biolegend or Miltenyi). In vitro, we investigated whether conditioned media from RPE cells treated under CEP+NHS sub-lytic MAC conditions would affect monocyte migration across a transwell as a proxy for understanding whether blockade of sub-lytic MAC signaling in RPE cells in vivo might reduce macrophage/microglia cell infiltration analogous to what we observed in the NaIO3-treated TLR2–/–mice. We observed increased cell migration across a transwell membrane toward the chamber with media transferred from RPE cells when compared with controls (Figure 6H), MCP-1 levels in the transferred media are shown for context (Figure 6I). We next assessed the effect of MCP-1 on CD86 as a proxy for monocyte activation. (J) CD86 MFI in CD45+CD66b-CD14+CD16+ monocytes treated with MCP-1 for 24 h. Experiments were carried out in triplicate and data are mean ± SE for three separate experiments.
Tags: anti-mouse CD86 (B7-2) 2D10; anti-mouse CD86 (B7-2) 2D10 mAb
4‐1BB costimulation promotes bystander activation of human CD8 T cells
Gschwendtner, A., et al. Eur J Immunol. 2021 May;51(5):1265-1276. doi: 10.1002/eji.202048863. PMID: 33626292
After blocking of the harvested cells from the coculture experiments for 20 min at 4°C with 20% AB‐serum in PBS, the cells were stained for 30 min with fluorophore‐labeled antibodies and pHLA‐A2 tetramers at 4°C. The following antibodies were used: CD80 PE (2D10), CD86 PE (IT2.2), 4‐1BBL PE (5F4), CD70 PE (113‐[16]), CD3 BV510 (UCHT1), CD4 BV421 (RPA‐T4), CD8 APC‐Cy7 (HIT8a), HLA‐A,B,C APC (W6/32), and CD56 PE‐Cy7 (CMSSB). CD8 T‐cells proliferating in response to cognate antigen express elevated levels of TIM‐3 and PD‐1. (A,B) The expression of various cell surface markers related to activation, costimulation, or coinhibition on proliferated CD8 T cells after stimulation with eAPC‐CD86 (A) or eAPC‐4‐1BBL (B) was assessed in 5 independent experiments using one donor per experiment. The percentages of marker positive cells within the pHLA‐A2 tetramer‐positive and ‐negative CD8+CFSElow T cells were compared. (C) Representative dot plots of these five independent experiments showing the expression of PD‐1 and TIM3 after stimulation with eAPC‐CD86 or eAPC‐4‐1BBL. Previously described Jurkat‐triple parameter reporter cells expressing 4‐1BB or control‐triple parameter reporter cells were cultured with or without T‐cell stimulator cells (TCS) at a ratio of 2.5:1. Urelumab (purchased from Creative Biolabs, Shirley, NY) was added at the indicated concentrations [50]. In additional experiments, these reporter cells were cocultured with K562‐HLA‐A2, K562‐HLA‐A2‐CD86, or K562‐HLA‐A2‐4‐1 BBL.
Tags: anti-mouse CD86 (B7-2) 2D10 antibody in vivo; anti-mouse CD86 (B7-2) 2D10 in animal model
Enzymatically-Processed Wheat Bran Enhances Macrophage Activity and Has in Vivo Anti-Inflammatory Effects in Mice
Kim, J. Y., et al. PLoS One. 2016 May 5;11(5):e0152979. doi: 10.1371/journal.pone.0152979. PMID: 27153047
Cells were blocked with rat anti-mouse CD16/CD32 (BD Pharmingen, San Diego, CA, USA) at 4 °C for 5 min and then stained for 30 min with FITC-conjugated anti-mouse SRA, anti-mouse CD11b, anti-CD40, PE-conjugated anti-mouse CD11b, anti-mouse CD36, and anti-mouse CD86 (all BD Pharmingen) on ice in the dark. CD86 is involved in the presentation of antigens to Th cells and is used as a marker for activated macrophages [27,28]. This molecule was not suppressed by the treatment of enzymatically-processed wheat bran, indicating that CD86 and CD40 are differentially regulated. We demonstrated that the anti-inflammatory effect of enzymatically-processed wheat bran occurred in response to systemic injection of LPS. As seen with the ex vivo data, the levels of pro-inflammatory cytokines TNF-α and IL-6 in serum were decreased. Regulation of contact dependent inflammatory molecules expressed by LPS-stimulated macrophages after oral administration of wheat bran. Cells isolated from each group were stimulated with LPS for 24 h. The cells were stained for FITC-conjugated CD40 antibody (ab) and PE-conjugated CD11b ab or FITC-conjugated CD11b ab and PE-conjugated CD86 ab.
Tags: anti-mouse CD86 (B7-2) 2D10 mAb in animal model; anti-mouse CD86 (B7-2) 2D10 in cancer research
Soluble CD80 Restores T Cell Activation and Overcomes Tumor Cell Programmed Death Ligand-1-mediated Immune Suppression
Hain, R., et al. J Immunother. 2013 Sep;36(7):386-95. doi: 10.1097/CJI.0b013e31829f9e6a. PMID: 23912944
Tumor cells were cultured in cover-glass slides containing 8 chambers (Thermo Scientific, Waltham, MA), washed with excess PBS containing 10% FCS, and labeled sequentially with mAb 5H1, anti-mouse IgG-Alexa Fluor 488 (Invitrogen, Grand Island, NY), and CD80-Alexa Fluor 647 (BioLegend, clone 2D10) (30min each step, with PBS/FCS washes after each antibody). CD28−/− splenocytes were suppressed by MELF10 cells and the suppression was not reversed by either CD80-Fc or CD86-Fc, indicating that costimulation through CD28 is integral to the ability of CD80-Fc to block PDL1-PD1 interactions. A potential drawback to using CD80-Fc as a reagent to inhibit PDL1-PD1 interactions is its potential to suppress T cell activation by binding to T cell-expressed CTLA4 (39, 40). In the experiments presented here, CD80-Fc increases, rather than decreases, T cell production of IFNγ (see figure 2), making it unlikely that CD80-Fc is interacting with CTLA4. (E) Splenocytes from CD28−/− mice were activated with PMA and ionomycin and co-cultured with MELF10 mouse tumor cells ± CD80-Fc, CD86-Fc, or TROY-Fc. Data are representative of 3, 3, 3, and 3 independent experiments for (A), (B), (C), and (D) respectively. (D) COS/PDL1/CD80 cells were plated in slides containing 8 individual chambers, stained with 5H1 + anti-mouse IgG-Alexa Fluor 488 and CD80-Alexa Fluor 647 (mAb 2D10), and analyzed by confocal microscopy. PDL1 is false colored green and CD80 is false colored red.
Tags: anti-mouse CD86 (B7-2) 2D10 mAb in cancer research; anti-mouse CD86 (B7-2) 2D10 mAb in mouse tumor model
B7-1 and B7-2 co-stimulatory molecules are required for mercury-induced autoimmunity
Gillespie, K. M., et al. Clin Exp Immunol. 2002 Feb;127(2):270-8. doi: 10.1046/j.1365-2249.2002.01789.x. PMID: 11882027
Mice were injected i.p. with 250 μg anti-B7-1 (CD80; clone 1G10) or anti-B7-2 (CD86; clone 2D10) every 3 days concurrently with HgCl2 treatment to neutralize costimulatory signals in vivo. B7-1 (CD80) and B7-2 (CD86) molecules on antigen presenting cells play important roles in providing co-stimulatory signals required for activation and expansion of autoreactive T cells. Moreover, some reports have suggested that these molecules may have distinct functions in the differentiation of Th1 and Th2 cells. Mercury-induced autoimmunity in H-2s mice is characterized by lymphoproliferation of T and B cells, serum increases in IgG1 and IgE and production of antinucleolar antibodies (ANoA). To examine the contributions of B7 co-stimulatory molecules to this model, susceptible mice were treated with antibodies to B7-1, B7-2, or both during the development of mercury-induced autoimmunity.
Tags: bioactivity of anti-mouse CD86 (B7-2) 2D10; anti-mouse CD86 (B7-2) 2D10 of low endotoxin
For more references about anti-mouse CD86 (B7-2) antibody (Clone: 2D10), please contact our scientific support team with message@sydlabs.com.