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Rat igg1κ

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Rat IgG1κ is an isotype control antibody derived from rat. It is designed to be used as a control for experiments involving rat antibodies.

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Lab products found in correlation

Nylon mesh filters, by Avantor (2 mentions) Gi254023x, by Merck Group (2 mentions) Live dead fixable aqua stain, by Thermo Fisher Scientific (2 mentions) Ifnγ cytokines, by Thermo Fisher Scientific (2 mentions) Cxcl16 antibody, by BD (2 mentions) Tnf α, by Thermo Fisher Scientific (2 mentions) Facscanto, by BD (2 mentions) Propidium iodide, by Thermo Fisher Scientific (1 mentions) Anti il 23p19, by Thermo Fisher Scientific (1 mentions) Cytofix cytoperm kit, by BD (1 mentions) Lsrfortessa cell analyzer, by BD (1 mentions) Polyplus, by Polyplus Transfection (1 mentions) Mirvana mirna inhibitor, by Thermo Fisher Scientific (1 mentions) Armenian hamster isotype control, by BioXCell (1 mentions) Rat igg2b, by Bio-Rad (1 mentions) Rat igg2aκ, by BioLegend (1 mentions) Clone mf1, by Bio-Rad (1 mentions)

5 protocols using rat igg1κ

1

Investigating CXCL16 Expression on Tumor and Mucosal Cells

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4T1 tumor cells were cultured for 24 hours in media containing 20 ng/mL TNFα and 20 ng/ml IFNγ cytokines (Peprotech) in the presence of 1 μM ADAM10 inhibitor to prevent membrane shedding (GI254023X, Sigma). Cells were dissociated with an enzyme-free dissociation buffer and homogenized to a single cell suspension. Mucosal epithelial cells were harvested directly from a tumor naive BALB/c mouse and were mechanically homogenized and filtered over 70 μm nylon mesh filters (VWR) to obtain a single cell suspension. Isolated cells were stained with a LIVE/DEAD Fixable Aqua stain (Thermo Fisher) and blocked with anti-mouse CD16/CD32 Fc Block (2.4G2) for 10 minutes prior to antibody staining. Cells were stained with either CXCL16 antibody (12–81, BD Biosciences) or isotype control (Rat IgG1κ, Biolegend). All data was acquired on a BD FacsCanto flow cytometer (BD Biosciences) and analyzed using FlowJo software (Treestar).
Christian L.S., Wang L., Lim B., Deng D., Wu H., Wang X.F, & Li Q.J. (2021). Resident memory T cells in tumor-distant tissues fortify against metastasis formation. Cell reports, 35(6), 109118.
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2

Investigating CXCL16 Expression on Tumor and Mucosal Cells

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4T1 tumor cells were cultured for 24 hours in media containing 20 ng/mL TNFα and 20 ng/ml IFNγ cytokines (Peprotech) in the presence of 1 μM ADAM10 inhibitor to prevent membrane shedding (GI254023X, Sigma). Cells were dissociated with an enzyme-free dissociation buffer and homogenized to a single cell suspension. Mucosal epithelial cells were harvested directly from a tumor naive BALB/c mouse and were mechanically homogenized and filtered over 70 μm nylon mesh filters (VWR) to obtain a single cell suspension. Isolated cells were stained with a LIVE/DEAD Fixable Aqua stain (Thermo Fisher) and blocked with anti-mouse CD16/CD32 Fc Block (2.4G2) for 10 minutes prior to antibody staining. Cells were stained with either CXCL16 antibody (12–81, BD Biosciences) or isotype control (Rat IgG1κ, Biolegend). All data was acquired on a BD FacsCanto flow cytometer (BD Biosciences) and analyzed using FlowJo software (Treestar).
Christian L.S., Wang L., Lim B., Deng D., Wu H., Wang X.F, & Li Q.J. (2021). Resident memory T cells in tumor-distant tissues fortify against metastasis formation. Cell reports, 35(6), 109118.
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3

Single-cell Immune Phenotyping and Cytokine Analysis

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Single-cell suspensions were pretreated with anti-CD16/32 antibody (clone: 2.4G2; TONBO Biosciences, San Diego, CA, USA) to block the Fc receptors. The cells were subsequently stained with the antibody mixture of CD45.2 (clone: 104; BioLegend, San Diego, CA, USA) and CD49f (clone: GoH3; eBioscience, San Diego, CA, USA). Propidium iodide (Invitrogen™, Thermo Fisher Scientific) was used to exclude the dead cells. For intracellular cytokine staining, the cells were stained with surface markers prior to fixation and permeabilization using Cytofix/Cytoperm kit (BD Biosciences, San Jose, CA, USA), according to the manufacturer’s protocol. Anti-IL-23p19 (clone: fc23cpg; eBiosciences) and RatIgG1κ (clone: RTK2071; BioLegend) were used as isotype controls. The stained samples were examined using LSRFortessa™ Cell Analyzer (BD Biosciences). The flow cytometry data were analyzed using FlowJo software (TreeStar, Ashland, OR, USA).
Park Y.J., Kim Y.H., Lee E.S, & Kim Y.C. (2022). Comparative Analysis of Single-Cell Transcriptome Data Reveals a Novel Role of Keratinocyte-Derived IL-23 in Psoriasis. Frontiers in Immunology, 13, 905239.
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4

Immunomodulation of Melanoma Progression

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WT and miR-146a-/- mice received 200μl intraperitoneal (i.p.) injections of Isotype control antibody (200μg/mouse /treatment) (Rat IgG1, κ; Biolegend, cat#400427) or IFN-γ blocking antibody (200μg/mouse/treatment) (R4.6A2, Biolegend, cat#505707) on day 0, 4, 8, and 12.
For translational in vivo experiments, WT mice injected with melanoma in the tail vein on day 0, received anti-PD-1 antibody, i.p. on days 1, 4 8, 16, and 22. Control mice were treated in parallel with an Armenian hamster isotype control purchased from BioXcell, (cat# BE0091). Oligonucleotides inhibiting miRNA-146a (ThermoFisher mirVana™ miRNA Inhibitor, Cat#: 4464088 ID: MH10722) or scramble controls (ThermoFisher mirVana™ miRNA Inhibitor, Negative Control #1, Cat#: 4464079) were administered via tail vein injections on days 5 and 9. In vivo-jetPEI® (Polyplus, cat# 201-50G), facilitated in vivo delivery of oligonucleotides across cell membranes, for 60μg of oligonucleotides injected in 200μl volume of 5% Glucose/in vivo-jet-PEI, per product protocol.
Mastroianni J., Stickel N., Andrlova H., Hanke K., Melchinger W., Duquesne S., Schmidt D., Falk M., Andrieux G., Pfeifer D., Dierbach H., Schmitt-Graeff A., Meiss F., Boerries M, & Zeiser R. (2018). miR-146a controls immune response in the melanoma microenvironment. Cancer research, 79(1), 183-195.
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5

Monoclonal Antibodies for Mouse Tetraspanins

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Monoclonal antibodies (mAb) against mouse tetraspanins were CD9 (Cat. No. MCA2749, clone MF1 Bio-Rad), CD63 (Cat. No. 143902, clone NVG-2, Biolegend), CD81 (Cat. No. MCA1846, clone Eat2 Bio-Rad), and matching isotype controls rat IgG2b (Bio-Rad), rat IgG2aκ (Biolegend) and hamster IgG1 (Bio-Rad), respectively. MAb to other cell surface molecules were CD36 (Cat. No. 102602, clone HM36), CD44 (Cat. No. 103002, clone IM7) CD47 (Cat. No. 127502, clone miap301) CD98 (Cat. No. 128202, clone RL388), CD172a (Cat. No. 144002, clone P84) (all from Biolegend), and DC-STAMP (Cat. No. MABF39, clone 1A2 Millipore). Isotype controls were Armenian hamster IgG, rat IgG2bκ, rat IgG2aκ, rat IgG2aκ, rat IgG1κ, (Biolegend), and IgG2aκ (Millipore), respectively. The secondary antibodies used for flow cytometry were anti-rat IgG-FITC (Cat. No. F-9387, Sigma) and anti-hamster IgG-FITC (Cat. No. MCA2357, Bio-Rad). All antibodies were used at saturating binding concentrations.
Elgawidi A., Mohsin M.I., Ali F., Watts A., Monk P.N., Thomas M.S, & Partridge L.J. (2020). A role for tetraspanin proteins in regulating fusion induced by Burkholderia thailandensis. Medical Microbiology and Immunology, 209(4), 473-487.
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