Isotype control 2a3

Manufactured by BioXCell

Sourced in Lebanon, United States

The Isotype control (2A3) is a laboratory reagent used in flow cytometry and other immunoassays. It serves as a negative control to establish baseline levels of non-specific binding. The 2A3 isotype control is a purified mouse immunoglobulin of the IgG2a isotype, which lacks reactivity to known antigens.

Automatically generated - may contain errors

Lab products found in correlation

11 protocols using isotype control 2a3

Adoptive Transfer and In Vitro Differentiation of TCR Transgenic T Cells

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Naïve T cells from TcR Tg mice were obtained from pooled spleen and lymph nodes as previously described (25 (link)). Resulting TcR Tg cells were routinely >97% TcR⁺ and expressed a characteristic naive phenotype (small size, CD62L^hi, CD44^lo and CD25^lo). T_H1 or T_c1 effectors were generated from naïve TcR Tg T cells cultured as previously described (25 (link)–27 (link)). In vitro-generated memory cells were obtained from effector cultures that were washed several times and rested for at least 3 days in media free of antigen (Ag) or exogenous cytokine. In some experiments, cells were labeled with carboxyfluorescein succinimidyl ester (CFSE) (Molecular Probes) prior to adoptive transfer. All cell populations were adoptively transferred to unprimed mice in 200 μl PBS by i.v. injection.
Mice were injected i.p. with IL-6-neutalizing Ab (MP5-20F3), or isotype control Ab (HRPN) (BioXcell) as indicated. In some experiments, mice were treated i.p. with 0.5 mg of anti Ly-6G Ab to deplete neutrophils (1A8) or isotype control (2A3) (BioXcell) on 0, 2, 4, and 6 dpi.

Strutt T.M., McKinstry K.K., Kuang Y., Finn C., Hwang J.H., Dhume K., Sell S, & Swain S.L. (2016). Direct IL-6 signals maximize protective secondary CD4 T cell responses against influenza. Journal of immunology (Baltimore, Md. : 1950), 197(8), 3260-3270.

+ Open protocol

+ Expand

Ly6G+ Cell Depletion Protocol

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Anti-Ly6G depleting antibody clone 1A8 or isotype control 2A3 (BioXcell, West Lebanon, NH) was administered as described by Nandi et al^{50 (link)}. Ly6G^Pos cell depletion was verified by FACS analysis of CD11b^High GR1^Pos Ly6C^Neg cell populations (Supplementary Fig. 2a).

Lovewell R.R., Baer C., Mishra B.B., Smith C.M, & Sassetti C.M. (2020). Granulocytes act as a niche for Mycobacterium tuberculosis growth. Mucosal immunology, 14(1), 229-241.

+ Open protocol

+ Expand

Immunization and Infection Protocols

Check if the same lab product or an alternative is used in the 5 most similar protocols

Influenza virus infections were performed intranasally (i.n) with 6,500 VFU of A/PR8/34 (PR8) in 100 μl of PBS. Acute LCMV infections were established by intraperitoneal injection with 2 × 10⁵ PFU LCMV-Armstrong. In some experiments mice were immunized with 50 μg of recombinant hemagglutinin (HA) adsorbed to Alum (Imject® Alum, Thermo Scientific™). BM chimeric mice were generated by irradiating the indicated recipient mice with 950 Rads from an X-ray source delivered in 2 equal doses administered 4–5 h apart. Following irradiation, mice were intravenously injected with 5 × 106 total BM cells and were allowed to reconstitute for 8–10 weeks before influenza infection. In indicated experiments, experimental animals received an intraperitoneal injection of 50 μg/kg of diphtheria toxin (DT - Sigma) at the indicated time points. In some experiments, mice were intraperitoneally administered recombinant IL-2 (National Cancer Institute) at the indicated time points. Control mice received injections of PBS. In some experiments mice were treated with 500 μg of a mix of anti-IL-2 neutralizing antibodies (JES6-1A12 and S4B6) or 500 μg of Isotype control (2A3), all obtained from BioXcell.

Botta D., Fuller M.J., Marquez-Lago T.T., Bachus H., Bradley J.E., Weinmann A.S., Zajac A.J., Randall T.D., Lund F.E., León B, & Ballesteros-Tato A. (2017). Dynamic regulation of T Follicular Regulatory cell responses by interleukin 2 during influenza infection. Nature immunology, 18(11), 1249-1260.

+ Open protocol

+ Expand

Neutrophil Depletion in Mouse RV1b Infection

Check if the same lab product or an alternative is used in the 5 most similar protocols

The experimental protocol is shown in Fig. 6e. Mice were injected i.p. with 500μg of neutrophil-depleting anti-Ly6G (α-Ly6G; 1A8; BioXCell, Lebanon, NH) or isotype control (2A3; BioXCell) antibodies in 200μl of PBS 24h before RV1b inoculation. Mice were euthanized 1 or 2 days after RV1b inoculation for BALF neutrophil and dsDNA analysis or NET detection, respectively (see below).

Toussaint M., Jackson D.J., Swieboda D., Guedán A., Tsourouktsoglou T.D., Ching Y.M., Radermecker C., Makrinioti H., Aniscenko J., Edwards M.R., Solari R., Farnir F., Papayannopoulos V., Bureau F., Marichal T, & Johnston S.L. (2017). Host DNA released by NETosis promotes rhinovirus-induced type 2 allergic asthma exacerbation. Nature medicine, 23(6), 681-691.

+ Open protocol

+ Expand

Tumor growth inhibition by ENT

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

BALB/c or BALB/c.scid mice were injected with 1 × 10⁵ TS/A or 4T1 cells into the mammary pad on day 0. The length and width of tumors were measured by caliper and tumor volume calculated by 0.4 × length × width². Tumor-bearing mice were treated daily with 20 mg/kg ENT (unless otherwise indicated) dissolved in 15% DMSO, 20% Kolliphor EL, 65% PBS or with the same volume of vehicle alone via intraperitoneal injection beginning on day 4 after tumor cell injection and continuing for two weeks unless otherwise stated. The chosen dose was based on that used in previous studies (22 , 23 (link)).
Depleting antibodies against CD4 (GK1.5), CD8 (2.43), CD25 (PC61.5.3), CD19 (1D3), B220 (RA3.3A1/6.1), anti-rat kappa (Mar 18.5) and isotype control (2A3) were obtained from BioXCell. For CD4, CD8 and CD25 depletion, 200 μg antibody was administered intraperitoneally on days −3, 1 and weekly intervals thereafter. For B cell depletion, 200 μg of anti-CD19 and anti-B220 was administered on day −10 and the anti-rat-kappa given on day −8. 200 μg anti-CD19 was then given on day −5, day −3, day 1 and weekly intervals thereafter. For combination experiments, 200 μg anti-PD1 (BioXCell, clone RMP1-14) or isotype antibody (BioXCell, clone 2A3) was administered intraperitoneally on days 10, 13, and 16.

McCaw T.R., Li M., Starenki D., Liu M., Cooper S.J., Arend R.C., Forero A., Buchsbaum D.J, & Randall T.D. (2019). Histone deacetylase inhibition promotes intratumoral CD8+ T cell responses, sensitizing murine breast tumors to anti-PD1. Cancer immunology, immunotherapy : CII, 68(12), 2081-2094.

+ Open protocol

+ Expand

Murine Immune Regulation Experiments

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

C57BL/6, B6.RAG1^−/−Tyrp1^B-w TRP-1 TCR transgenic, and B6.OPG^−/− mice were purchased from The Jackson Laboratory. Mice were treated intraperitoneally with 100 µg anti-RANKL (IK22/5) or isotype control (2A3; Bio X Cell) antibody three times per week as stated in the text. B6.OT-II, B6.RIP-mOVA, and B6.IRBP^−/− mice were described previously (Anderson et al., 2005 (link); DeVoss et al., 2006 (link)). Mice were treated at 3–5 wk of age and harvested at time points indicated in the text. All mice were housed and bred in specific pathogen-free conditions in animal facilities at UCSF or UNC, Chapel Hill. Animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) at UCSF or UNC, Chapel Hill.

Khan I.S., Mouchess M.L., Zhu M.L., Conley B., Fasano K.J., Hou Y., Fong L., Su M.A, & Anderson M.S. (2014). Enhancement of an anti-tumor immune response by transient blockade of central T cell tolerance. The Journal of Experimental Medicine, 211(5), 761-768.

+ Open protocol

+ Expand

Anti-inflammatory Treatments in Disease

Check if the same lab product or an alternative is used in the 5 most similar protocols

All antibody treatments were given intraperitoneally (i.p.) in a 200-μl volume. A total of 150 μg anti-Ly6G (clone 1A8) or isotype control (2A3) (BioXCell) was given every 24 h from 4 to 12 dpi. A total of 500 μg anti-TNF-α (XT3.11) or isotype control (HRPN) (BioXCell) was given on 5 and 7 dpi. A total of 150 μg anti-TRAIL (Cambridge Bioscience) or vehicle (PBS) was given either (continuous treatment) every 48 h from 1 to 9 dpi, (early treatment) at 1 and 3 dpi or (late treatment) at 6 and 8 dpi.

Ellis G.T., Davidson S., Crotta S., Branzk N., Papayannopoulos V, & Wack A. (2015). TRAIL+ monocytes and monocyte-related cells cause lung damage and thereby increase susceptibility to influenza–Streptococcus pneumoniae coinfection. EMBO Reports, 16(9), 1203-1218.

+ Open protocol

+ Expand

Immunomodulatory Reagents and Antineoplastic Drugs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mouse recombinant GM-CSF, mouse recombinant IL-6, human recombinant GM-CSF, and human recombinant G-CSF were purchased from Miltenyi Biotec (Bologna, Italy). K^b-restricted OVA_257–₂₆₄ peptide (SIINFEKL), was synthesized by JPT (Berlin, Germany). Necrostatin-1, 5Z-7-Oxozeaenol (253863) and TPCA-1 (507475-17-4) were purchased from Sigma-Aldrich (Saint Louis, MO, USA) and pan caspase Inhibitor (Z-VAD-FMK) from R&D Systems (Minneapolis, MN, USA). InVivoMAb anti-mouse PD-1 (RMP1-14) and isotype control (2A3) were purchased from Bioxcell (Lebanon, NH, USA). Chemotherapeutic drugs: 5-fluorouracil (51-21-8); gemcitabine (95058-81-4); docetaxel (114977-28-5); paclitaxel (33069-62-4); oxaliplatin (61825-94-3); cisplatin (15663-27-1), etoposide (33419-42-0), irinotecan (100286-90-6), fludarabine (21679-14-1) and carboplatin (41575-94-4) were purchased by Cayman Chemical (Ann Arbor, MI, USA).

Fiore A., Ugel S., De Sanctis F., Sandri S., Fracasso G., Trovato R., Sartoris S., Solito S., Mandruzzato S., Vascotto F., Hippen K.L., Mondanelli G., Grohmann U., Piro G., Carbone C., Melisi D., Lawlor R.T., Scarpa A., Lamolinara A., Iezzi M., Fassan M., Bicciato S., Blazar B.R., Sahin U., Murray P.J, & Bronte V. (2018). Induction of immunosuppressive functions and NF-κB by FLIP in monocytes. Nature Communications, 9, 5193.

+ Open protocol

+ Expand

Ly6G+ Cell Depletion Protocol

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lovewell R.R., Baer C., Mishra B.B., Smith C.M, & Sassetti C.M. (2020). Granulocytes act as a niche for Mycobacterium tuberculosis growth. Mucosal immunology, 14(1), 229-241.

+ Open protocol

+ Expand

Immunotherapies for Cancer Treatment

Check if the same lab product or an alternative is used in the 5 most similar protocols

Isotype control (2A3, catalogue number BE0089), anti-PD-L1 (clone 10 F.9G2, catalogue number BE0101) and anti-CTLA-4 (9H10, catalogue number BE0131) were purchased from BioXcell. Mice were treated every 4 days for up to 2 doses. Where indicated mice were treated at days 14, 16, 18 and 20 post-tumor inoculation with 25 µg FTY720 (Sigma). Anti-asialo GM-1 (catalogue number 986-10001, FUJIFILM Wako Pure Chemical Corporation) and anti-CD8α (clone YTS 169.4, catalogue number BE0117, BioXcell) were used for in vivo NK cell and CD8⁺ T cell depletion, respectively. Anti-CD3 (clone 145-2C11) and anti-CD28 (clone 37.51) used to stimulate murine T cells were obtained from BD Pharmingen. Cytokines used for the stimulation of murine or Human immune cells were purchased from Biolegend.

Todd K.L., Lai J., Sek K., Huang Y.K., Newman D.M., Derrick E.B., Koay H.F., Nguyen D., Hoang T.X., Petley E.V., Chan C.W., Munoz I., House I.G., Lee J.N., Kim J.S., Li J., Tong J., N. de Menezes M., Scheffler C.M., Yap K.M., Chen A.X., Dunbar P.A., Haugen B., Parish I.A., Johnstone R.W., Darcy P.K, & Beavis P.A. (2023). A2AR eGFP reporter mouse enables elucidation of A2AR expression dynamics during anti-tumor immune responses. Nature Communications, 14, 6990.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!