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Mouse igg1

Manufactured by Southern Biotech
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Sourced in Italy

Mouse IgG1 is a type of immunoglobulin G (IgG) antibody isolated from mouse serum or cell cultures. IgG1 is the most abundant subclass of IgG antibodies and plays a crucial role in the adaptive immune response.

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Lab products found in correlation

Anti c myc mab clone 9e10, by Merck Group (3 mentions) Pe conjugated goat anti mouse igg, by Southern Biotech (3 mentions) Facscalibur flow cytometer, by BD (2 mentions) Cellquest, by BD (1 mentions) Facs fortessa, by BD (1 mentions) Intracellular fixation and permeabilization buffer, by Thermo Fisher Scientific (1 mentions) α cd3 pe, by BD (1 mentions) α cd11b fitc, by BioLegend (1 mentions) α ifnγ apc, by BD (1 mentions) Jam c, by R&D Systems (1 mentions) Mouse igg2a, by Southern Biotech (1 mentions) Mouse igg2b, by Southern Biotech (1 mentions) Ps9 gsk3β, by Cell Signaling Technology (1 mentions) Py705 stat3, by Cell Signaling Technology (1 mentions) Alexa conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Hrp conjugated secondary antibody, by Southern Biotech (1 mentions) Stat3, by Cell Signaling Technology (1 mentions) Gp130, by Santa Cruz Biotechnology (1 mentions) Filgotinib, by Selleck Chemicals (1 mentions) Ps727 stat3, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

Goat anti-mouse IgG1-HRP (26 citations) Biotinylated goat-anti-mouse IgG1 (4 citations) Mouse IgG1 isotype control (15H6, (3 citations) Goat anti-mouse IgG1-biotin (3 citations) Goat anti-mouse IgG/IgG1/IgG2a/IgG2b/IgG3 antibodies labeled with Horseradish peroxidase (3 citations) Mouse anti-human IgG1 Fc-HRP (3 citations) Anti-mouse IgG1-APC (2 citations) Goat anti-mouse IgG1 antibody conjugated with horseradish peroxidase (2 citations) Goat anti-mouse IgG1 antibody (2 citations) Goat Anti-Mouse IgG1-AP (2 citations)

9 protocols using mouse igg1

1

Comprehensive Immune Profiling by FACS

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Cell suspensions were stained for FACS analysis with the following antibodies: α-CD3-PE (BD Biosciences), α-CD8-BV605 (BD Biosciences), α-CD45-PercP (BD Biosciences), α-CD45-PE (BD Biosciences), α-CD11b-FITC (BioLegend), α-CD11c-FITC (BioLegend), α-CTLA4-PE (BD Biosciences), α-granzyme B-Alexa fluor 647 (BD Biosciences), α-IFNγ-APC (BD Biosciences), α-KI67-BV421 (BD Biosciences), α-TIM3-PE (eBioscience), α-TIGIT-PE (BD Biosciences), α- TNFα-PE (BD Biosciences), α-IL17r-PE (BD Biosciences), and the corresponding isotype control (mouse IgG1, Southern Biotech). The live/dead fixable near-infrared dye (Invitrogen) was used to exclude dead cells. Intracellular fixation and permeabilization buffers from eBioscience were used for cytokine staining.
Acquisition was performed on FACS BD Fortessa and the dedicated software CellQuest (BD Biosciences). Data was analyzed with FlowJo 7.5.5 software (TreeStar Inc).
D’Amico L., Menzel U., Prummer M., Müller P., Buchi M., Kashyap A., Haessler U., Yermanos A., Gébleux R., Briendl M., Hell T., Wolter F.I., Beerli R.R., Truxova I., Radek Š., Vlajnic T., Grawunder U., Reddy S, & Zippelius A. (2019). A novel anti-HER2 anthracycline-based antibody-drug conjugate induces adaptive anti-tumor immunity and potentiates PD-1 blockade in breast cancer. Journal for Immunotherapy of Cancer, 7, 16.
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2

Antibody Detection and Signaling Pathways

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The following primary antibodies were used (dilution 1:1,000 for Western blot, 1:200 for immunofluorescence and flow cytometry): GAPDH (mouse SC-25778; Santa Cruz Biotechnology), GP130 (rabbit SC-656 for Western blot, mouse SC-376280 for immunofluorescence; Santa Cruz Biotechnology; and mouse ab34324 for flow cytometry; Abcam), APLNR (mouse FAB8561R and MAB856; R&D Systems), JAMC (mouse FAB11891A; R&D Systems), STAT3 (rabbit 9132; Cell Signaling), pS727-STAT3 (rabbit 9134; Cell Signaling), pY705-STAT3 (rabbit 9145; Cell Signaling), pS235/S236-S6 (rabbit 2211 for Western blot and rabbit 5316 for flow cytometry; Cell Signaling), pS9-GSK3β (rabbit 9336; Cell Signaling), ELMOD1 (rabbit ab127541; Abcam), RAB5 (rabbit ab218624; Abcam) and RAB7 (rabbit 9367; Cell Signaling). HRP-conjugated secondary antibodies (anti-rabbit, mouse Ig, mouse IgG1, mouse IgG2a, and mouse IgG2b) were purchased from Southern Biotech. Alexa-conjugated secondary antibodies were from Life Technologies. FLAER (Alexa Fluor 488–conjugated proaerolysin) was purchased from Cedarlane, MBCD was from Sigma-Aldrich, and Stattic, ruxolitinib, and filgotinib were all from Selleckchem. [Pyr1]-Apelin-13 was resuspended and aliquoted according to the supplier’s recommendation (Phoenix).
Trillet K., Jacobs K.A., André-Grégoire G., Thys A., Maghe C., Cruard J., Minvielle S., Diest S.G., Montagnac G., Bidère N, & Gavard J. (2021). The glycoprotein GP130 governs the surface presentation of the G protein–coupled receptor APLNR. The Journal of Cell Biology, 220(9), e202004114.
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3

Phenotypic Characterization of CAR T Cells

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All experiments were carried out on a flow cytometer FACSCalibur (BD Bioscience), and data obtained were analyzed using FlowJo software (TreeStar). CAR expression was evaluated through anti-c-myc mAb (clone 9E10; Sigma-Aldrich) or the relative isotype control (mouse IgG1, Southern Biotech, Milan, Italy), followed by a secondary antibody (PE-conjugated goat anti-mouse IgG; Southern Biotech). T cell phenotype was evaluated using mAb to CD62L, CCR7, CD27, CD28, CD57, PD-1, TIM-3, LAG-3 (all from eBioscience), Annexin V and FasL (from BD Bioscience), and the relative isotype controls purchased from the same companies. All FACS plots presenting CAR T cell phenotype data were conducted on gated CAR+ cells.
Zuccolotto G., Penna A., Fracasso G., Carpanese D., Montagner I.M., Dalla Santa S, & Rosato A. (2021). PSMA-Specific CAR-Engineered T Cells for Prostate Cancer: CD28 Outperforms Combined CD28-4-1BB “Super-Stimulation”. Frontiers in Oncology, 11, 708073.
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4

CTLA-4 Blockade Assay with Flow Cytometry

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The blocking ability of the anti-CTLA-4 mAb was confirmed using flow cytometry with CD80-Ig or CD86-Ig and CTLA-4-EGFP expressing CHO-DG44 cells. CTLA-4 expressing cells were incubated in PBS containing 10% goat serum12 at room temperature for 15 min. After pretreatment, the cells were incubated with different concentrations of anti-CTLA-4 mAb (1.25, 2.5, 5.0, 10, and 20 μg/ml) for 20 min at 25 °C. Mouse IgG1 (Southern Biotech) was used as an isotype antibody. The cells were washed twice, and 0.2 μg/ml CD80-Ig or CD86-Ig was then added. After incubating for 20 min at 25 °C, the cells were washed twice. CD80-Ig or CD86-Ig was detected using flow cytometry with Alexa Fluor 647-conjugated anti-rabbit IgG (H + L) goat IgG (Thermo Fisher Scientific).
Watari K., Konnai S., Maekawa N., Okagawa T., Suzuki Y., Murata S, & Ohashi K. (2019). Immune inhibitory function of bovine CTLA-4 and the effects of its blockade in IFN-γ production. BMC Veterinary Research, 15, 380.
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5

Chicken PBMC Immune Cell Profiling

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The PBMCs of immunized chickens were collected, and the diversity of CD4+/CD3+ and CD8+/CD3+ detected. Briefly, 5×106 PBMCs were double labelled with two antibodies: an aliquot of cells was incubated with mouse anti-chicken CD4-PE (phycoerythrin, PE) (Southern Biotech; Cat No.: 8255-09) and anti-chicken CD3- fluorescein isothiocyanate, FITC) (Southern Biotech; Cat No.: 8200-02), and another aliquot of cells was incubated with anti-chicken CD8α-PE (Southern Biotech; Cat No.: 8405-09) and anti-chicken CD3-FITC (Southern Biotech; Cat No.: 8200-02). Fluorescent-activated cell sorting (FACS) controls were produced by incubating PBMCs with mouse Ig G1 (Southern Biotech) conjugated to PE and FITC. Cells were washed twice with PBST buffer after a 30 min incubation at room temperature, then suspended in PBS buffer and analyzed by FACS (BD Biosciences, Franklin Lakes, NJ, USA).
Xu H., Li L., Li R., Guo Z., Lin M., Lu Y., Hou J., Govinden R., Deng B, & Chenia H.Y. (2022). Evaluation of dendritic cell-targeting T7 phages as a vehicle to deliver avian influenza virus H5 DNA vaccine in SPF chickens. Frontiers in Immunology, 13, 1063129.
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6

Blocking CTLA-4 Binding to CD80/CD86

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The ability of Boch4C2 to block the binding of bovine CTLA-4 to CD80/CD86 was assessed using an ELISA. MaxiSorp Immuno Plates (Thermo Fisher Scientific) were coated with 1 μg/ml CD80-Ig or CD86-Ig in PBS for 30 min at 37°C and then blocked with PBS containing 1% bovine serum albumin (Sigma-Aldrich) for 30 min at 37°C. Biotin-conjugated CTLA-4-Ig was preincubated with Boch4C2, bovine IgG (Sigma-Aldrich), 4C2-D9, or mouse IgG1 (Southern Biotech) at molar ratios (CTLA-4-Ig:antibody)=1:0, 1:0.1, 1:0.5, 1:1, 1:2, 1:5, and 1:10 for 30 min at 37°C. These reagents were added to the microplate and incubated for 30 min at 37°C. Binding of biotin-conjugated CTLA-4-Ig to CD80-Ig or CD86-Ig was detected using the NeutrAvidin Horseradish Peroxidase Conjugate (Thermo Fisher Scientific) for 30 min at 37°C. TMB One Component Substrate (Bethyl Laboratories, Montgomery, TX, USA) was then added to the microplate, and the reaction was stopped using 0.18 M H2SO4. Absorbance at 450 nm was measured using an MTP-900 microplate reader (Corona Electric, Hitachinaka, Japan). Three duplicate independent experiments were performed. The relative binding of CTLA-4-Ig to CD80-Ig or CD86-Ig was calculated from the absorbance of samples preincubated with each antibody, and the absorbance of the sample preincubated without antibody, which was defined as 100%.
WATARI K., KONNAI S., OKAGAWA T., MAEKAWA N., SAJIKI Y., KATO Y., SUZUKI Y., MURATA S, & OHASHI K. (2021). Enhancement of interleukin-2 production by bovine peripheral blood mononuclear cells treated with the combination of anti-programmed death-ligand 1 and cytotoxic T lymphocyte antigen 4 chimeric monoclonal antibodies. The Journal of Veterinary Medical Science, 84(1), 6-15.
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7

Establishment of Anti-Bovine CTLA-4 mAb

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Establishment of anti-bovine CTLA-4 mouse mAb was outsourced to Cell Engineering Corporation (Osaka, Japan). The reactivity of polyclonal antibodies from hybridomas was screened using ELISA. Bovine CTLA-4 monoclonal antibody was selected by flow cytometry using CTLA-4-EGFP expressing CHO-DG44 cells as described above. An anti-bovine CTLA-4 monoclonal antibody (4G2-A3) was selected by the screening and purified for this study. The specificity of the monoclonal antibody to CTLA-4 was confirmed using flow cytometry. In brief, CTLA-4 or CD28 expressing cells were incubated in PBS containing 10% goat serum (Sigma-Aldrich) at room temperature for 15 min to suppress nonspecific binding to the Fc receptor. After pretreatment, the cells were incubated with 10 μg/ml anti-CTLA-4 mAb or a control Ab (mouse IgG1, Southern Biotech) for 20 min at room temperature. The cells were washed twice, and anti-CTLA-4 mAb was detected with Alexa Fluor 647-conjugated anti-mouse IgG (H + L) F (ab’)2 (Thermo Fisher Scientific).
Watari K., Konnai S., Maekawa N., Okagawa T., Suzuki Y., Murata S, & Ohashi K. (2019). Immune inhibitory function of bovine CTLA-4 and the effects of its blockade in IFN-γ production. BMC Veterinary Research, 15, 380.
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8

Immunophenotyping of CAR-expressing Cells

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CAR-expressing cells were labeled with the anti-c-myc mAb (clone 9E10; Sigma-Aldrich) or the isotype control (mouse IgG1, Southern Biotech, Milan, Italy), followed by a secondary antibody (PE-conjugated goat anti-mouse IgG; Southern Biotech). Cell surface markers were labeled using APC-, FITC- or PE-conjugated antibodies to CD4, CD8, CD57, CD27, CD28, CD62L (BioLegend, San Diego, CA, USA), CCR7 (eBioscience, San Diego, CA, USA), and the relative isotype controls purchased from the same companies.
Zuccolotto G., Fracasso G., Merlo A., Montagner I.M., Rondina M., Bobisse S., Figini M., Cingarlini S., Colombatti M., Zanovello P, & Rosato A. (2014). PSMA-Specific CAR-Engineered T Cells Eradicate Disseminated Prostate Cancer in Preclinical Models. PLoS ONE, 9(10), e109427.
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9

CAR-expressing Cells Immunophenotyping

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CAR-expressing cells were labeled with the anti-c-myc mAb (clone 9E10; Sigma-Aldrich) or the isotype control (mouse IgG1; Southern Biotech), followed by a secondary antibody (PE-conjugated goat anti-mouse IgG; Southern Biotech). Cell surface markers were labeled using PE-conjugated antibodies to CD85j, NKG2D, CD158d, CD27, and with the relative isotype controls (BioLegend). Flow cytometry analyses were conducted using a FACSCalibur flow cytometer and FlowJO software (BD Biosciences).
Montagner I.M., Penna A., Fracasso G., Carpanese D., Dalla Pietà A., Barbieri V., Zuccolotto G, & Rosato A. (2020). Anti-PSMA CAR-Engineered NK-92 Cells: An Off-the-Shelf Cell Therapy for Prostate Cancer. Cells, 9(6), 1382.
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