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Rat igg2b isotype control

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The Rat IgG2b isotype control is a laboratory reagent used as a negative control in immunoassays and flow cytometry experiments. It is a purified immunoglobulin G2b (IgG2b) antibody from rat, which serves as a non-specific binding control to determine background or non-specific signal in experiments involving rat-derived antibodies.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Dta 1, by BioXCell (2 mentions) Rat igg1 isotype control, by BioXCell (2 mentions) Anti cd8 antibody, by BioXCell (2 mentions) Sulindac, by Merck Group (1 mentions) Colo205, by American Type Culture Collection (1 mentions) Nf κb inhibitor bay 11 7082, by Merck Group (1 mentions) Cell culture medium, by Thermo Fisher Scientific (1 mentions) Anti mouse pd l1 antibody, by BioXCell (1 mentions) Ht 29, by American Type Culture Collection (1 mentions) Caco 2, by American Type Culture Collection (1 mentions) Anti cd8 mab 2, by BioXCell (1 mentions) Anti t bet and biotin conjugated anti b220 antibodies, by BioLegend (1 mentions) Rat igg1, by BioXCell (1 mentions) Rmil 33, by BioLegend (1 mentions) Rm il 7, by BioXCell (1 mentions) Anti cd4 antibody clone gk1, by BioXCell (1 mentions) Pe cy7 hamster anti mouse cd11c, by BioLegend (1 mentions) Bv421 rat anti mouse mhc 2, by BioLegend (1 mentions) Antibodies against mouse cd3, by BioLegend (1 mentions)

Close spelling variants of this product

Rat IgG2b (38 citations) Isotype control (29 citations) IgG2b (19 citations) IgG2b isotype control (6 citations) IgG2b isotype (3 citations) Rat IgG2b isotype (2 citations) Rat IgG2a and IgG2b isotype controls (2 citations)

28 protocols using rat igg2b isotype control

1

Depletion of CD90+ ILCs in Mice

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Depletion of CD90+ ILCs was performed as previously described, with some modifications (36 (link)). Two hundred and fifty micrograms of a mAb to CD90.2 or an isotype control rat IgG2b (BioXCell, West Lebanon, NH) was given by means of intra-peritoneal injection a total of three times at 3-day intervals. Intestinal ECs and LP cells were collected 2 days after the final injection.
Goto Y., Obata T., Kunisawa J., Sato S., Ivanov I.I., Lamichhane A., Takeyama N., Kamioka M., Sakamoto M., Matsuki T., Setoyama H., Imaoka A., Uematsu S., Akira S., Domino S.E., Kulig P., Becher B., Renauld J.C., Sasakawa C., Umesaki Y., Benno Y, & Kiyono H. (2014). Innate lymphoid cells regulate intestinal epithelial cell glycosylation. Science (New York, N.Y.), 345(6202), 1254009.
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2

Colorectal Cancer Cell Line Characterization

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The murine colon cancer cell line CT26 and the human colon cancer cell lines, Colo205, HT29, and Caco-2, were purchased from ATCC (Manassas VA, USA). We authenticate our cell lines at regular intervals of 12 months in addition to two authentications at the beginning and end of the project using the services provided by IDEXX BioAnalytics (Columbia, MO). The authentication includes short tandem repeat (STR) profiling, mycoplasma testing, and cross-species contamination checking. In this project, only early passages of the cell lines (<10) were utilized. Cell culture mediums were purchased from Thermo Fisher Scientific (Carlsbad CA, USA) and used for cell culture after mixing with 10% fetal bovine serum (Thermo Fisher Scientific). Cells were cultured at 37°C and in a 5% CO2 humidified incubator. Sulindac, CMC (Carboxymethylcellulose), and NF-κB inhibitor (Bay11-7082) were purchased from Sigma-Aldrich (St Louis MO, USA). CMC was used as the vehicle of Sulindac for in vivo study. Mouse anti-PD-L1 antibody and isotype control rat IgG2b were purchased from BioXcell (West Lebanon NH, USA). Sulindac sulfide was purchased from Alfa Aesar (Haverhill MA, USA).
Yi B., Cheng H., Wyczechowska D., Yu Q., Li L., Ochoa A.C., Riker A.I, & Xi Y. (2021). Sulindac modulates the response of proficient MMR colorectal cancer to anti-PD-L1 immunotherapy. Molecular cancer therapeutics, 20(7), 1295-1304.
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3

CD4+ and CD8+ T Cell Depletion

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CD8+ and CD4+ T cells were depleted in mice as previously described [22 (link)]. Briefly, 0.5 mg of anti-CD8 mAb 2.43 (BioXCell) and 0.35 mg of anti-CD4 mAb 1.5 (BioXCell), or 0.85 mg of isotype control rat IgG2b (BioXCell) was iv-injected into mice 24 hours prior to parasite challenge. T cell depletion was confirmed before each challenge by collecting 50–100 μl of peripheral blood via the retro-orbital plexus from each mouse and assaying peripheral blood lymphocytes by flow cytometry staining for CD19, CD3, CD4 and CD8.
Sack B.K., Keitany G.J., Vaughan A.M., Miller J.L., Wang R, & Kappe S.H. (2015). Mechanisms of Stage-Transcending Protection Following Immunization of Mice with Late Liver Stage-Arresting Genetically Attenuated Malaria Parasites. PLoS Pathogens, 11(5), e1004855.
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4

Immune Checkpoint Inhibition and T-cell Analysis

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Purified monoclonal anti-mouse PD-L1 (10F.9G2), anti-mouse IFN-γ (XMG1.2), and isotype control rat IgG2b and rat IgG1 were purchased from BioXCell. Recombinant mouse (rm) IFN-γ was purchased from Peprotech. The muscarinic acetycholine receptor (M3R) peptide was synthesized by Biomatik Corporation. For Flow cytometry, anti-CD4, anti-CD8 and anti-CD44 antibodies were purchased from BioLegend. For immunohistological chemistry, biotin conjugated anti-CD4 antibody was obtained from eBioscience, anti-T-bet and biotin conjugated anti-B220 antibodies were from BioLegend, and biotin conjugated anti-rat IgG2bwere purchased from Vector Laboratories. For immunofluorescence staining, anti-aquaporin 5 (AQP5) and Alexa Fluor647-conjugated anti-rabbit IgG antibodies were purchased from Abcam.
Zhou J., Jin J.O., Kawai T, & Yu Q. (2016). Endogenous programmed death ligand-1 restrains the development and onset of Sjӧgren’s syndrome in non-obese diabetic mice. Scientific Reports, 6, 39105.
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5

Isolation and Stimulation of Murine VAT ILC2s

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Murine VAT and human peripheral ILC2s were isolated to >95% purity using the FACS Aria III cell sorter. For in vivo stimulation of murine VAT ILC2s, carrier free rm-IL-33 (Biolegend, San Diego, CA, 1 µg/mouse in 200 µL) was administered intraperitoneally to mice on three consecutive days. On day 4, murine ILC2s were isolated based on the lack of expression of classical lineage markers (CD3ε, CD45R, Gr-1, CD11c, CD11b, Ter119, TCRγδ, and FCεRI) and expression of CD45, ST2, and CD117. Isolated ILC2s were stimulated (5 × 104/mL) with rm-IL-2 (10 ng/mL) and rm-IL-7 (10 ng/mL) for 48 h at 37 °C in presence of GITR agonist DTA-1 (5 µg/mL) from BioXCell, West Lebanon, New Hampshire (BE0063) or the monoclonal antibody rat IgG2b isotype control (BioXCell). For adoptive transfer experiments, 2.5 × 105 purified ILC2s were adoptively transferred intravenously in 200 µL PBS into the recipients at the start of the indicated treatment.
Galle-Treger L., Sankaranarayanan I., Hurrell B.P., Howard E., Lo R., Maazi H., Lewis G., Banie H., Epstein A.L., Hu P., Rehan V.K., Gilliland F.D., Allayee H., Soroosh P., Sharpe A.H, & Akbari O. (2019). Costimulation of type-2 innate lymphoid cells by GITR promotes effector function and ameliorates type 2 diabetes. Nature Communications, 10, 713.
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6

High-fat diet and GITR agonist in mice

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When indicated, mice were fed a high fat diet (HFD, Rodent diet with 60 kcal% Fat, D12492i) from Research Diets Inc. (New Brunswick, New Jersey) for the indicated times, as described before40 (link). All other mice were fed a normal chow diet (NCD). For in vivo experiments investigating the effect of GITR engagement, GITR agonist DTA-1 (1 mg/mouse, BioXCell, West Lebanon, New Hampshire, BE0063), or the monoclonal antibody rat IgG2b isotype control (BioXCell) (1 mg/mouse) was administered intraperitoneally every 4 days from the indicated start of treatment until termination of the experiment.
Galle-Treger L., Sankaranarayanan I., Hurrell B.P., Howard E., Lo R., Maazi H., Lewis G., Banie H., Epstein A.L., Hu P., Rehan V.K., Gilliland F.D., Allayee H., Soroosh P., Sharpe A.H, & Akbari O. (2019). Costimulation of type-2 innate lymphoid cells by GITR promotes effector function and ameliorates type 2 diabetes. Nature Communications, 10, 713.
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7

CD4+ T Cell Depletion in Mice

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Mice were injected intraperitoneally with 500 µg of anti-CD4 antibody (clone GK1.5, BioXCell) or rat IgG2b isotype control (BioXCell) on day 21 and again on day 22 post infection. Mice were analyzed 7 days after antibody administration. anti-CD4 (clonse RMA4.4) was used to monitor the efficiency of T cell depletion by flow cytometry.
Kirchner F.R, & LeibundGut-Landmann S. (2020). Tissue-resident memory Th17 cells maintain stable fungal commensalism in the oral mucosa. Mucosal Immunology, 14(2), 455-467.
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8

C1498 Mouse Model of AML with Anti-PD-L1

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This study was approved by the “Wenzhou Medical University Animal Care and Use Committee” and carried out in accordance with the recommendations of “institutional guidelines, Wenzhou Medical University Animal Care and Use Committee.” The C1498 mouse model of AML was established as described previously with minor modifications. Briefly, exponentially growing C1498 cells (5 × 106) were resuspended in 100 μl PBS, and subsequently intravenously injected into the tail vein of recipient mice, which had been already exposed to 5 Gy myeloablative irradiation 4 h before. According to the methods previously reported (23 (link)), these mice were administrated intraperitoneally with 7.5 mg/kg anti-mouse PD-L1 antibody (clone: 10F.9G2) or with rat IgG2b isotype control (both from BioXcell, West Lebanon, NH, USA) on days 0, 3, 6, 9, 12, and sacrificed on day 15.
Dong Y., Han Y., Huang Y., Jiang S., Huang Z., Chen R., Yu Z., Yu K, & Zhang S. (2020). PD-L1 Is Expressed and Promotes the Expansion of Regulatory T Cells in Acute Myeloid Leukemia. Frontiers in Immunology, 11, 1710.
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9

Anti-Ly6G Depletion in Mice

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Mice were treated i.p. with 500 µg anti-Ly6G (Gr-1) (RB6-8C5, Bio X Cell, West Lebanon, NH) or rat IgG2b isotype control (Bio X Cell) every other day from the day of infection for 8 days.
Kamdar K., Khakpour S., Chen J., Leone V., Brulc J., Mangatu T., Antonopoulos D.A., Chang E.B., Kahn S.A., Kirschner B.S., Young G, & DePaolo R.W. (2016). Genetic and metabolic signals during acute enteric bacterial infection alter the microbiota and drive progression to chronic inflammatory disease. Cell host & microbe, 19(1), 21-31.
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10

Tracking DC Migration and VCAM-1 Blocking

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Typically, 1–1.5 million CMFDA- or DR-labeled LPS mature DCs (ratio 1:1) were injected in INF or CTR mouse ears or footpads. Auricular and popliteal LNs were harvested at different time points, passaged through a 40-µm cell strainer and stained with BV421 rat anti-mouse MHC-II (BioLegend) and Pe/Cy7 hamster anti-mouse CD11c (BioLegend). Cell suspensions were acquired on a CytoFlex S and the total number of migratory labeled (CMFDA or DR) DCs analyzed in FlowJo. For VCAM-1 blocking experiments in CHS inflammation, 0.5 mg of VCAM-1 blocking antibody was administered i.p. after oxazolone challenge and 8 h before the adoptive transfer. 24 h after challenge, 1:1 ratios of LPS mature tln1−/− and WT DCs were injected together with 25 µg VCAM-1 blocking antibody (6C7.1; Engelhardt et al., 1998 (link)) or rat IgG1 isotype control (BioXCell) in INF ears and footpads. For integrin α4 blocking experiments, 1:1 ratios of LPS mature tln1−/− and WT DCs were coinjected with 10 µg integrin α4 blocking antibody (PS/2; Miyake et al., 1991 (link); BioXCell) or rat IgG2b isotype control (BioXCell) in either INF ears or footpads. 14 h (footpads) and 17 h (ear skin) later, dLNs were harvested and processed as described above. The whole LN was acquired on a Cytoflex S, and the total number of migratory labeled (CMFDA or DR) DCs analyzed in FlowJo.
Arasa J., Collado-Diaz V., Kritikos I., Medina-Sanchez J.D., Friess M.C., Sigmund E.C., Schineis P., Hunter M.C., Tacconi C., Paterson N., Nagasawa T., Kiefer F., Makinen T., Detmar M., Moser M., Lämmermann T, & Halin C. (2021). Upregulation of VCAM-1 in lymphatic collectors supports dendritic cell entry and rapid migration to lymph nodes in inflammation. The Journal of Experimental Medicine, 218(7), e20201413.
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