Rat igg1

Manufactured by BioXCell

Sourced in United States

Rat IgG1 is a type of immunoglobulin G (IgG) antibody derived from rat species. It is a widely used laboratory reagent for various immunological applications.

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20 protocols using rat igg1

Modulating Wound Healing in Mice

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Mouse model of Vγ1D was implemented by intraperitoneal injection of 200 μg anti-Vγ1 Ab (clone 2.11; BioXcell, USA) 3 days before wound excision. Vγ4D was generated by anti-Vγ4 Ab (clone UC3-10A6; BioXcell, USA). Control mice were intraperitoneally administrated isotype control (armenian hamster IgG) (BioXcell, USA). Neutralization mouse models were conducted by injecting subcutaneously into wound margin 0, 1, and 2 days following excision with 20 μg/wound of anti-IGF-1 Ab (R&D Systems, Minneapolis, MN, USA), anti-IL-17A Ab (clone 17F3; BioXcell, USA), anti-IFN-γ Ab (clone XMG1.2; BioXcell, USA), anti-IL-1β Ab (clone B122; BioXcell, USA), anti-IL-23 Ab (clone BE0051; BioXcell, USA), anti-CCL20 Ab (clone 114906; R&D Systems, USA), respectively. IL-17A neutralization of low and high doses was performed by 2 and 200 μg/wound of anti-IL-17A Ab, respectively. Isotype control mice received equivalent doses of isotype Abs subcutaneously. Isotype control Abs of IGF-1, IL-17A, IL-1β, IL-23, CCL20 are poly goat IgG, mouse IgG1, armenian hamster IgG, rat IgG2, and rat IgG1 (BioXcell), respectively. Mouse models of recombinant cytokine addition were generated by subcutaneously injecting rIGF-1/rIL-17A/rIL-1β/rIL-23 (R&D Systems, Minneapolis, MN, USA) with the doses of 2, 20, 200 ng/wound, respectively, in the wound margin. Control mice were administrated with sterile PBS.

Li Y., Wang Y., Zhou L., Liu M., Liang G., Yan R., Jiang Y., Hao J., Zhang X., Hu X., Huang Y., Wang R., Yin Z., Wu J., Luo G, & He W. (2018). Vγ4 T Cells Inhibit the Pro-healing Functions of Dendritic Epidermal T Cells to Delay Skin Wound Closure Through IL-17A. Frontiers in Immunology, 9, 240.

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Treg Cell Ablation and Depletion Protocol

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For Treg cell ablation studies, DT (Sigma-Aldrich) was injected
intraperitoneally (i.p.) at 50 µg per kg of body weight at the indicated
times. For neutralization and depletion studies, anti-mouse IFN-γ (1 mg,
clone XMG1.2, Bio X Cell) or anti-mouse CD11c (500 µg, clone N418, Bio X
Cell) or anti-mouse CD8 (250 µg, clone 53–6.72, Bio X Cell) or
anti-mouse CD4 (150 µg, clone GK1.5, Bio X Cell) or anti-mouse CD25 (500
µg, clone PC-61.5.3, Bio X Cell) were injected i.p. at the indicated
times. Isotype control antibodies used were Rat IgG1 or Armenian hamster IgG or
Rat IgG2a from Bio X Cell, respectively.

Jang J.E., Hajdu C.H., Liot C., Miller G., Dustin M.L, & Bar-Sagi D. (2017). Crosstalk between Regulatory T Cells and Tumor-Associated Dendritic Cells Negates Anti-tumor Immunity in Pancreatic Cancer. Cell Reports, 20(3), 558-571.

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Neutralizing Antibodies for Mouse Cytokines

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Neutralizing antibodies to mouse TNF (rat anti-mouse; clone XT3.11) and IL-17A (mouse anti-mouse; clone 17F3) were in vivo grade from Bio X Cell (Lebanon, NH, USA). Mouse anti-mouse TWEAK neutralizing antibody (clone mP2D10) was produced by Biogen, Inc. Isotype control antibodies were rat IgG1, mouse IgG1, and mouse IgG2a, all from Bio X Cell (Lebanon, NH, USA) for the TNF, IL-17A and TWEAK neutralizing antibodies, respectively. All recombinant human cytokines, TWEAK, TNF, and IL-17A, were from R&D Systems (Minneapolis, MN, USA).

Gupta R.K., Gracias D.T., Figueroa D.S., Miki H., Miller J., Fung K., Ay F., Burkly L, & Croft M. (2021). TWEAK Functions with TNF and IL-17 on Keratinocytes and is a Potential Target for Psoriasis Therapy. Science immunology, 6(65), eabi8823.

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T Cell and NK Cell Depletion

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To deplete NK cells, CD4⁺ T cells and CD8⁺ T cells, tumour-bearing mice were intraperitoneally injected with anti-NK1.1 (clone PK136, BioXCell), anti-CD4 (clone GK1.5, BioXCell), anti-CD8-α (clone 2.43, BioXCell) or isotype control (RatIgG1,BioXcell) antibodies at an initial dose of 400 mg 1 d before treatment, followed by 200 mg every 3 d. Depletion of CD8⁺ T cells, CD4⁺ T cells and NK cells was confirmed using flow cytometry analysis of peripheral blood mononuclear cells.

Wang F., Su H., Xu D., Dai W., Zhang W., Wang Z., Anderson C.F., Zheng M., Oh R., Wan F, & Cui H. (2020). Tumour sensitization via the extended intratumoural release of a STING agonist and camptothecin from a self-assembled hydrogel. Nature biomedical engineering, 4(11), 1090-1101.

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Modulating Cytokine Responses via Blocking Antibodies

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Specific blocking antibodies to TNF-α and IL-6 were used to elucidate the functional importance of modulation of these cytokines in co-culture. Blocking antibodies against TNF-α (XT3.11), IL-6 (MP5-20 F3) and isotype (Rat IgG1) control (all from Bio X Cell, NH, USA) were reconstituted in 1X PBS to 1.0 mg/ml. The antibodies were then serially diluted in culture media to obtain working stock concentrations of 20.0, 2.0 and 0.2 μg/ml. Cells were plated either in 12-well plates (for Griess assays) or in 96-well plates (for proliferation assays) as described earlier. Cells were stimulated with 1 μg/ml LPS after overnight incubation. Equal volumes of diluted antibody were added to cultures to obtain a final concentration of 10.0, 1.0 and 0.1 μg/ml. Culture supernatants were collected at 24 and 48 hours and assayed for NO production using Griess assay. Proliferation was analysed at 48 hours by pulsing the cultures with ³H-TdR for the final 6 hours of incubation.

Jose S., Tan S.W., Ooi Y.Y., Ramasamy R, & Vidyadaran S. (2014). Mesenchymal stem cells exert anti-proliferative effect on lipopolysaccharide-stimulated BV2 microglia by reducing tumour necrosis factor-α levels. Journal of Neuroinflammation, 11, 149.

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Treg Cell Ablation and Depletion Protocols

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For Treg cell ablation studies, DT (Sigma-Aldrich) was injected intraperitoneally (i.p.) at 50 μg per kg of body weight at the indicated times. For neutralization and depletion studies, anti-mouse IFN-γ (1 mg, clone XMG1.2, Bio X Cell), anti-mouse CD11c (500 μg, clone N418, Bio X Cell), anti-mouse CD8 (250 μg, clone 53-6.72, Bio X Cell), anti-mouse CD4 (150 μg, clone GK1.5, Bio X Cell), or anti-mouse CD25 (500 μg, clone PC-61.5.3, Bio X Cell) was injected i.p. at the indicated times. Isotype control antibodies used were rat IgG1, Armenian hamster IgG, or rat IgG2a from Bio X Cell, respectively.

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Antibody-Mediated T Cell Depletion and Treg Abrogation

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The following antibodies were given i.p. at 200µg per injection: anti-CD8α (Clone: 2.43; BioXcell), anti- CD8β (Clone: 53-5.8; BioXcell), anti-CD25 (Clone: PC61; eBioscience), as well as rat IgG2B (Clone: LTF-2; BioXcell) and rat IgG1 (Clone: HRPN; BioXcell) isotype controls. For in vivo T_Reg cell depletion with Diphteria Toxin (DT; Sigma-Aldrich), Foxp3^DTR mice were given 50µg/kg of DT i.p. every other day as previously described (28 (link),29 (link)).

Rivas M.N., Lee Y., Wakita D., Chiba N., Dagvadorj J., Shimada K., Chen S., Fishbein M.C., Lehman T.J., Crother T.R, & Arditi M. (2017). CD8+ T cells contribute to the development of coronary arteritis in the Lactobacillus casei extract-induced murine model of Kawasaki Disease. Arthritis & rheumatology (Hoboken, N.J.), 69(2), 410-421.

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Anti-IFNAR-1 Antibody Modulates Mtb Infection

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For some experiments, mice were treated with anti-IFNAR-1 antibodies. One month after control and alcohol diet feeding, the mice were challenged with aerosolized Mtb. After 3 months, the mice received 0.3 mg of anti-IFNAR-1 (BioXcell, Clone: MAR1-5A3, Catalog number: BP0241) or isotype-matched control Ab (rat IgG1, Clone: MOPC-21, Catalog number: BE0083) intravenously every 4 days for up to 2 months.

Tripathi D., Welch E., Cheekatla S.S., Radhakrishnan R.K., Venkatasubramanian S., Paidipally P., Van A., Samten B., Devalraju K.P., Neela V.S., Valluri V.L., Mason C., Nelson S, & Vankayalapati R. (2018). Alcohol enhances type 1 interferon-α production and mortality in young mice infected with Mycobacterium tuberculosis. PLoS Pathogens, 14(8), e1007174.

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Blocking Antibody Treatment in Mice

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The blocking antibodies anti-mMCAM (clone 15) (described in [22 (link)]) and α4 integrin neutralizing antibody (clone: PS/2, BioXCell, New Hampshire, USA) as well as the appropriate isotype control antibodies (rat IgG1, clone: HRPN and rat IgG2b, clone: LTF-2; both BioXCell) were used at a concentration of 10 mg/kg body weight. Mice were treated every other day with i.p. injections of the respective antibody from the indicated day on.

Breuer J., Korpos E., Hannocks M.J., Schneider-Hohendorf T., Song J., Zondler L., Herich S., Flanagan K., Korn T., Zarbock A., Kuhlmann T., Sorokin L., Wiendl H, & Schwab N. (2018). Blockade of MCAM/CD146 impedes CNS infiltration of T cells over the choroid plexus. Journal of Neuroinflammation, 15, 236.

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Concanavalin A and α-GalCer Induced Liver Injury

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Tumor-free littermates or mice bearing subcutaneous tumors were challenged with 12.5 μg/g type IV Concanavalin A (Con A) (Sigma, USA) or 2 μg/mouse α-Galactosylceramide (α-GalCer, Funakoshi LTD, Japan). Alanine/aspartate aminotransferase (ALT/AST) levels were determined in mouse sera. In some experiments, mice were fed with a butylated hydroxyanisole (BHA, Sigma, USA)-containing diet for 3 days (7 mg/g bodyweight) to block ROS production [41 (link)],[45 (link)],[49 (link)]. IFN-γ serum levels after Con A injection were quantified by ELISA (eBioscience, USA). Endogenous IFN-γ was blocked by treating mice with 200 μg rat-anti mouse anti IFN-γ antibody (clone XMG1.2, BioXCell, USA). The same dose of rat IgG1 (BioXCell, USA) was used as a control. For visualization of ROS production, mice were injected with 5 μg luminescent probe L-012 (Wako, USA) which shows luminescence after chemical reaction with ROS and imaged with IVIS Spectrum (Xenogen, USA). Bioluminescence was calculated based on photon flux (photon counts/second) obtained 5 min after injection, divided by liver area using Living Image software (Perkin Elmer, USA). Mice were sacrificed at the indicated time points. All animal studies were approved by the National Cancer Institute Bethesda Animal Care and Use Committee.

Kapanadze T., Medina-Echeverz J., Gamrekelashvili J., Weiss J.M., Wiltrout R.H., Kapoor V., Hawk N., Terabe M., Berzofsky J.A., Manns M.P., Wang E., Marincola F.M., Korangy F, & Greten T.F. (2015). CD40 dependent exacerbation of immune mediated hepatitis by hepatic CD11b+ Gr-1+ myeloid derived suppressor cells in tumor bearing mice. European journal of immunology, 45(4), 1148-1158.

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