Human CXCL4/PF4 Antibody (170138)

Detection of kfpxvaa9ffykrjoxefaf.jpg by Flow Cytometry

(a) Size-exclusion chromatograms. The elution profiles of lysozyme (14.4 kDa) and myoglobin (17 kDa) in blue and OHD4–12 (15.9 kDa) in orange are shown. (b) WB analysis in non-reduced (left bands, N.R.) and reduced (right bands, R.) conditions demonstrates the presence of OHD4–12 obtained following the mixing of CXCL4-S26C and CXCL12-L29C mutants in the presence of Cu2+ detected with anti-CXCL4 ( alpha CXCL4) or anti-CXCL12 ( alpha CXCL12) antibodies (see also Supplementary Fig. S1). (c) NMR spectroscopic analysis of OHD4–12 folding state. The 15N-HSQC NMR spectrum of the uniformly 15N-labeled 68 μM OHD4–12 in 90% H2O/10% D2O at pH 6.9 in the presence of 20 mM NaCl, collected at 40 °C. (d–f) Expansions from the OHD4–12 spectrum overlaid with CXCL4wt (blue) and CXCL12wt (red) 15N-HSQC NMR spectra. Known signal assignments for CXCL4 and CXCL12 are indicated. The 15N-HSQC NMR spectrum of the uniformly 15N-labeled 150 μM CXCL4 in 90% H2O/10% D2O at pH 5.0 in the presence of 20 mM NaCl was collected at 40 °C. The 15N-HSQC NMR spectrum of the uniformly 15N-labeled 51 μM CXCL12 in 20 MES buffer prepared with 90% H2O/10% D2O at pH 6.8 was collected at 25 °C. The difference in experimental conditions was due to the difference in solubility properties of CXCL4 and CXCL12. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36229490), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of CXCL4/PF4 by Western Blot

(a) Size-exclusion chromatograms. The elution profiles of lysozyme (14.4 kDa) and myoglobin (17 kDa) in blue and OHD4–12 (15.9 kDa) in orange are shown. (b) WB analysis in non-reduced (left bands, N.R.) and reduced (right bands, R.) conditions demonstrates the presence of OHD4–12 obtained following the mixing of CXCL4-S26C and CXCL12-L29C mutants in the presence of Cu2+ detected with anti-CXCL4 ( alpha CXCL4) or anti-CXCL12 ( alpha CXCL12) antibodies (see also Supplementary Fig. S1). (c) NMR spectroscopic analysis of OHD4–12 folding state. The 15N-HSQC NMR spectrum of the uniformly 15N-labeled 68 μM OHD4–12 in 90% H2O/10% D2O at pH 6.9 in the presence of 20 mM NaCl, collected at 40 °C. (d–f) Expansions from the OHD4–12 spectrum overlaid with CXCL4wt (blue) and CXCL12wt (red) 15N-HSQC NMR spectra. Known signal assignments for CXCL4 and CXCL12 are indicated. The 15N-HSQC NMR spectrum of the uniformly 15N-labeled 150 μM CXCL4 in 90% H2O/10% D2O at pH 5.0 in the presence of 20 mM NaCl was collected at 40 °C. The 15N-HSQC NMR spectrum of the uniformly 15N-labeled 51 μM CXCL12 in 20 MES buffer prepared with 90% H2O/10% D2O at pH 6.8 was collected at 25 °C. The difference in experimental conditions was due to the difference in solubility properties of CXCL4 and CXCL12. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36229490), licensed under a CC-BY license. Not internally tested by R&D Systems.