Cyclosporine (BioVision, Milpitas, CA, catalog no. 1522-1G), dosed at 0.75 mg suspended in 10 μl of dimethyl sulfoxide (Corning, catalog no. 25-950-CQC) and 90 μl of olive oil, was injected subcutaneously at the base of the mouse neck. The protocol was adapted from the publication of Klaus and Kunkl (25 (link)), but the dosing was halved to reduce the mouse fatality rate. Mice were treated with 250 μg of L243 (pan–HLA-DR, mouse IgG2a) or isotype control (mouse IgG2a, BioXCell, catalog no. BE0085) antibody in 100 μl of PBS biweekly by intraperitoneal injection (31 (link)). L243 (pan–HLA-DR) was a gift from J. Moon, Massachusetts General Hospital/Harvard Medical School, Boston, MA, USA. Mice receiving IL-2 (BioLegend, catalog no. 589104) treatment were administered 80,000 U/kg suspended in PBS by intraperitoneal injection. Mice were treated with UVB (302 to 312 nm) (100 mJ/cm2) three times weekly for up to 42 weeks using a UVP Blak-Ray Lamp UVB (VWR, Radnor, PA, catalog no. 36575-052) to simulate 25 to 50 min of sun exposure in Florida at midday in the summer (61 (link)). Mice receiving basiliximab were administered a single injection of 50 μg suspended in 100 μl of PBS intradermally and were monitored for at least 90 days after injection.
Mouse igg2a
Manufactured by BioXCell
Sourced in United States
Mouse IgG2a is a type of immunoglobulin G (IgG) antibody derived from mouse. It is a laboratory reagent used in various immunological research and assays.
Automatically generated - may contain errors
Lab products found in correlation
Mouse igg1, by BioXCell (3 mentions)
Interleukin 2 (il 2), by BioLegend (1 mentions)
Tumor necrosis factor (tnf), by R&D Systems (1 mentions)
Tweak, by R&D Systems (1 mentions)
Il 17a, by BioXCell (1 mentions)
Il 17a, by R&D Systems (1 mentions)
Rat igg1, by BioXCell (1 mentions)
Mouse cd4 t cell isolation kit, by Miltenyi Biotec (1 mentions)
Complete freund adjuvant (cfa), by Merck Group (1 mentions)
96 well half area plate, by Corning (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Tween 20, by Merck Group (1 mentions)
Goat serum, by Merck Group (1 mentions)
Skim milk, by Merck Group (1 mentions)
Synergy 2 plate reader, by Agilent Technologies (1 mentions)
Tmb substrate, by Thermo Fisher Scientific (1 mentions)
Rat igg2a, by BioLegend (1 mentions)
α nk1, by BioXCell (1 mentions)
Asgm1, by Fujifilm (1 mentions)
Clone ltf 2, by BioXCell (1 mentions)
17 protocols using mouse igg2a
1
Immunosuppressant and UV Treatments in Mice
2
Neutralizing Antibodies for Mouse Cytokines
3
Adoptive Transfer of Transgenic T and B Cells
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Adoptive transfers were performed as described previously (Smith et al., 2000 (link)). Briefly, single cell suspensions of LNs and spleens were prepared from OT-II and MD4 mice. The proportion of Tg B and T cells was determined by flow cytometry and 2–3 × 106 Tg B cells and 2–3 × 106 Tg T cells adoptively transferred to age-matched C57BL/6J recipients via tail vein injection. For imaging studies, DsRed expressing CD4+ OT-II T cells were magnetically sorted from OT-II × hCD2-DsRed animals using mouse CD4+ T cell isolation kits (Miltenyi Biotec, Surrey, UK). Mice were immunized via the footpad with 130 µg of OVA-HEL conjugate alone, with CFA (Sigma, Dorset, UK), or covalently bound to carboxylate-modified microspheres. Injection with 50 μl of PBS was used as a vehicle control. Popliteal LNs were excised for ex vivo study. In MHCII blocking experiments, 500 μg Y3P (BioXcell, New Hampshire, USA) or isotype control (mouse IgG2a; BioXcell) was given via tail vein injection. In T cell/DC interaction studies, LNs were imaged 2 hr after antibody treatment.
4
Humanized Anti-C1s Monoclonal Antibodies
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TNT009, a humanized anti‐C1s mAb (IgG4 isotype, stock solution: 1 mg/mL), and its parental mouse variant, TNT003 (IgG2a isotype, stock solution: 1 mg/mL), were obtained from True North Therapeutics, Inc. (South San Francisco, CA). Nonspecific human IgG4 (Abcam PLC, Cambridge, UK) or mouse IgG2a (Bio X Cell, West Lebanon, NH) served as the isotype controls. The targets of TNT009/TNT003 and other complement inhibitors used in this study are illustrated in Figure S1 .
5
Checkpoint Blockade Therapy Evaluation
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Anti-PD-1 (murine IgG1, clone 4H2) (20 (link)), anti-CTLA-4 non-depleting (ND; murine IgG1-D265A; a non-FcγR binding mutant with deficient FcγR-mediated depletion) and anti-CTLA-4 depleting (D; murine IgG2a; with competent FcγR-mediated depletion) (21 (link)). Antibody treatment was administered via intra-peritoneal (ip) injection at a dose of 10 mg/kg body weight every 3 days for a total of 3 doses. Mouse IgG2a (clone: C1.18.4; BioXCell, West Lebanon, NH) was used as an isotype control.
6
TYRP1 Binding Assay Protocol
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96-well half area plates (Corning) were coated with 1µg/mL recombinant human His-tagged TYRP1 (Sino Biologicals, Cat# 13224-H08H) overnight at 4°C. The following day, plates were blocked with 1x PBS containing 5% skim milk (Sigma), 10% goat serum (Milipore), 1% bovine serum albumin (BSA; Sigma), 1% fetal bovine serum (FBS, Lampire) and 0.2% Tween-20 (Sigma) for 2 hours at RT. Plates were subsequently incubated with serially diluted mouse sera or recombinant protein antibody-cytokine chimera (depending on assay) for 2 hours at 37°C before incubation with 1:20000 HRP-conjugated anti-mouse IgG H+L (Bethyl, Cat# A90-116P) for 1 hour at room temperature. In addition, mouse IgG2a (BioXcell, Catalog: C1.18.4) was used as an isotype control. Following this, plates were developed with TMB substrate (Thermo) for approximately 5 minutes at RT before being stopped with 2N H2SO4. Plates were read with the BioTEK Synergy 2 plate reader and absorbance measured at 450 and 570nm.
7
ICOS Blockade and ILC Depletion Protocol
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For ICOS blockade experiments, an anti-ICOSL antibody (#107410; BioLegend; 20 µg/20 µl in Sal) was delivered intranasally for three consecutive days 3 d before analysis in week 8. An isotype rat IgG2a (#400544; BioLegend) antibody was used as a control. For NK1.1 and ILCs cell depletion, we used as anti-NK1.1(#BE0036, clone PK136; Bio X Cell) and anti-CD90.2 (#BE0066, clone 30H12; Bio X Cell), respectively. Mouse IgG2a (#BE0085, clone C1.18.4; Bio X Cell) and rat IgG2b (#BE0090, clone LTF-2; Bio X Cell) antibodies were used as isotype controls, respectively. For ILC2 depletion in vivo, we treated mice with SR3335 (200 µg/dose, i.p.) or vehicle (ethanol diluted with Sal; final ethanol concentration 20 µl/dose) on two consecutive days before the recall challenge.
8
NK Cell Depletion in Ischemia-Reperfusion Injury
9
In vivo Murine Klebsiella pneumoniae Infection
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For infection experiments, mice were i.n. challenged with 5 × 104 CFUs of K. pneumoniae in 30 μl of sterile PBS. For in vivo antibody-mediated blocking experiments, mice were intraperitoneally (i.p.) injected with the following antibodies 24–48 h before infection: αMR1 (150 μg/mouse; clone 26.5; Biolegend, Mouse IgG2a), αIFNAR1 (200 μg/mouse; clone MAR1-5A3; BioXCell, Mouse IgG1), αSiglecH (150 μg/mouse; clone 440c; BioXCell, Rat IgG2b), or their respective isotype controls: Mouse IgG2a (150 μg/mouse; clone MOPC-173; Biolegend), Mouse IgG1 (200 μg/mouse; clone MOPC-21; BioXCell), and Rat IgG2b (150 μg/mouse; clone LTF-2; BioXCell). When indicated, MAIT cell expansion in vivo was performed through a repeated i.n. inoculation (3×) of 5-OP-RU (100 μM) and LPS (17.4 μg/mouse; Invivogen).
Bacterial loads were determined by counting CFUs after plating 100-fold dilution series of tissue homogenates obtained from bacteria-infected mice. Colonies were counted at 24 h.
Bacterial loads were determined by counting CFUs after plating 100-fold dilution series of tissue homogenates obtained from bacteria-infected mice. Colonies were counted at 24 h.
10
Influenza Infection Survival in Depleted Mice
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Mice (C57BL/6, female) were immunized three times at 2-week intervals. The mice were infected with 10 LD50 of PR8 on day 42 after initial immunization. To deplete immune cells, the mice were intraperitoneally injected with 450 μg of anti-CD4 (rat IgG2b; Bio X Cell, USA), anti-CD8 (rat IgG2b; Bio X Cell), anti-NK1.1 (mouse IgG2a; Bio X Cell), or isotype control antibody (mouse IgG2a; Bio X Cell) on days 39, 41, and 43. After virus infections, body weight and survival rate were monitored for 14 days.
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