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Rabbit anti rat ig

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Rabbit anti-rat Ig is a secondary antibody product that binds to rat immunoglobulins (Ig). It is produced by immunizing rabbits with rat Ig and purifying the resulting antibodies. This product can be used in various immunological techniques, such as Western blotting, ELISA, and immunohistochemistry, to detect and quantify rat Ig in samples.

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Anti-rabbit Ig (3 citations)

3 protocols using rabbit anti rat ig

1

T Cell Activation Signaling Assay

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Short-term–expanded CD4+ T cells (100 × 106) from WT and CD5OST mice or long-term–expanded WT, LATOST, and CD6OST CD4+ T cells (100 × 106) were incubated with anti-CD3 (0.2 µg per 106 cells; 145-2C11; Exbio) and anti-CD4 (0.2 µg per 106 cells; GK1.5; Exbio) antibodies on ice for 15 min, followed by one round of washing at 4°C. Cells were then incubated at 37°C for 5 min and then stimulated at 37°C with a purified rabbit anti-rat Ig (0.4 µg per 106 cells; Jackson ImmunoResearch) for 30, 120, and 300 s or left unstimulated. Stimulation was stopped by the addition of a twice-concentrated lysis buffer (100 mM Tris, pH 7.5, 270 mM NaCl, 1 mM EDTA, 20% glycerol, and 0.4% n-dodecyl-β-D-maltoside) supplemented with protease and phosphatase inhibitors. After 10 min of incubation on ice, cell lysates were centrifuged at 21,000 g for 5 min at 4°C. Postnuclear lysates were then used for affinity purification.
Mori D., Grégoire C., Voisinne G., Celis-Gutierrez J., Aussel R., Girard L., Camus M., Marcellin M., Argenty J., Burlet-Schiltz O., Fiore F., Gonzalez de Peredo A., Malissen M., Roncagalli R, & Malissen B. (2020). The T cell CD6 receptor operates a multitask signalosome with opposite functions in T cell activation. The Journal of Experimental Medicine, 218(2), e20201011.
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2

Temporal Dynamics of TCR Signaling

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Short-term expanded CD4+ T cells (100 × 106) from wild-type mice and OST-tagged mice were incubated with anti-CD3 (0.2 μg per 106 cells; 145-2C11, Exbio) and anti-CD4 (0.2 μg per 106 cells; GK1.5, Exbio) on ice for 15 min, followed by one round of washing at 4°C. Cells were then incubated at 37°C for 5 min and then stimulated at 37°C with a purified rabbit anti-rat Ig (0.4 μg per 106 cells; Jackson Immuno-Research) for 30, 120, 300, and 600 s or left unstimulated. Stimulation was stopped by the addition of a twice-concentrated lysis buffer (100 mM Tris, pH 7.5, 270 mM NaCl, 1 mM EDTA, 20% glycerol, 0.4% n-dodecyl-β-D-maltoside) supplemented with protease and phosphatase inhibitors. After 10 min of incubation on ice, cell lysates were centrifuged at 21,000 g for 5 min at 4°C. Post-nuclear lysates were then used for affinity purification.
Voisinne G., Kersse K., Chaoui K., Lu L., Chaix J., Zhang L., Menoita M.G., Girard L., Ounoughene Y., Wang H., Burlet-Schiltz O., Luche H., Fiore F., Malissen M., de Peredo A.G., Liang Y., Roncagalli R, & Malissen B. (2019). Quantitative interactomics in primary T cells unveils TCR signal diversification extent and dynamics. Nature immunology, 20(11), 1530-1541.
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3

Temporal Dynamics of TCR Signaling

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Short-term expanded CD4+ T cells (100 × 106) from wild-type mice and OST-tagged mice were incubated with anti-CD3 (0.2 μg per 106 cells; 145-2C11, Exbio) and anti-CD4 (0.2 μg per 106 cells; GK1.5, Exbio) on ice for 15 min, followed by one round of washing at 4°C. Cells were then incubated at 37°C for 5 min and then stimulated at 37°C with a purified rabbit anti-rat Ig (0.4 μg per 106 cells; Jackson Immuno-Research) for 30, 120, 300, and 600 s or left unstimulated. Stimulation was stopped by the addition of a twice-concentrated lysis buffer (100 mM Tris, pH 7.5, 270 mM NaCl, 1 mM EDTA, 20% glycerol, 0.4% n-dodecyl-β-D-maltoside) supplemented with protease and phosphatase inhibitors. After 10 min of incubation on ice, cell lysates were centrifuged at 21,000 g for 5 min at 4°C. Post-nuclear lysates were then used for affinity purification.
Voisinne G., Kersse K., Chaoui K., Lu L., Chaix J., Zhang L., Menoita M.G., Girard L., Ounoughene Y., Wang H., Burlet-Schiltz O., Luche H., Fiore F., Malissen M., de Peredo A.G., Liang Y., Roncagalli R, & Malissen B. (2019). Quantitative interactomics in primary T cells unveils TCR signal diversification extent and dynamics. Nature immunology, 20(11), 1530-1541.
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