Reagents were purchased from Invitrogen (Paisley, UK), unless stated otherwise. Recombinant human CXCL10 and CXCL11 were purchased from PeproTech EC, Ltd. (London, UK). The monoclonal mouse anti-haemagglutinin (HA) anti-HA.11 antibody was from Covance (Berkeley, CA, USA) and its corresponding IgG1 isotype control antibody from Sigma-Aldrich (Poole, UK). The anti-CXCR3 mAb (Clone 49801) was from R&D Systems (Abingdon, UK). The murine pre-B cell line L1.2 was maintained as described previously (Vaidehi et al., 2009 (link)). TAK-779 was obtained from the Programme EVA Centre for AIDS Reagents, NIBSC, UK, supported by the EC FP6/7 Europrise Network of Excellence, AVIP and NGIN consortia and the Bill and Melinda Gates GHRC-CAVD Project and was donated by Dr R. Gallo, University of Maryland School of Medicine. The synthesis of VUF 10085 has previously been described (Flier et al., 2001 (link); Storelli et al., 2007 (link)).
Igg1 isotype control antibody
Manufactured by Merck Group
Sourced in United States
The IgG1 isotype control antibody is a laboratory reagent used as a control in immunoassays and flow cytometry experiments. It is designed to have the same isotype as the primary antibody being used, but does not recognize any specific antigen. This allows researchers to distinguish between specific binding of the primary antibody and non-specific background signal.
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Lab products found in correlation
2 protocols using igg1 isotype control antibody
1
Chemokine Receptor CXCR3 Signaling Assay
2
RNA-binding protein SRSF1 immunoprecipitation
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After trypsinization, A549 cells were incubated with nuclear isolation buffer (1.28 M sucrose; 40 mM Tris pH 7.4; 20 mM MgCl 2 ; 4% Triton X-100). Nuclear pellets were resuspended in RIP buffer (150 mM KCl; 25 mM Tris pH 7.4; 5 mM EDTA; 0.5% NP40, 0.5 mM DTT; and protease and RNase inhibitors). After centrifugation, supernatants were split into two fractions and 10% was saved as input. Twenty micrograms of anti-SRSF1 antibody (Zymed) or 20 µg of IgG1 isotype control antibody (Sigma) were added to each fraction and incubated overnight at 4 °C in continuous rotation. Magnetic beads were added and incubated for 1 h at 4 °C in continuous rotation. Five washes with RIP buffer and two washes with PBS were performed to wash off unbound material. Pellets were resuspended in trizol reagent (Invitrogen). RNA was extracted according to the manufacturer's protocol. RT-PCR was performed by using the total amount of RNA extracted. RNA quantification was represented as percentage of input.
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