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Recombinant human sdf 1α

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Recombinant human SDF-1α is a protein produced in E. coli cells. It is a chemokine that plays a role in the migration and recruitment of cells.

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Lab products found in correlation

Igf1r, by Santa Cruz Biotechnology (1 mentions) Recombinant human igf 1, by Thermo Fisher Scientific (1 mentions) Fitc conjugated cholera toxin b subunit ctxb, by Merck Group (1 mentions) Cytofix fixation buffer, by BD (1 mentions) Methyl β cyclodextrin mβcd, by Merck Group (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) Mitotracker red cm h2xros, by Thermo Fisher Scientific (1 mentions) Mrs1754, by R&D Systems (1 mentions) 5 bdbd, by R&D Systems (1 mentions) Anti human cd4 allophycocyanin clone okt4, by BioLegend (1 mentions) Anti human cd69 fitc clone fn50, by BioLegend (1 mentions) Mouse anti human cd3 clone hit3a, by BD (1 mentions) Nf546, by R&D Systems (1 mentions) Rhod 2 am, by Thermo Fisher Scientific (1 mentions) Nf340, by R&D Systems (1 mentions) Fluo 4 am, by Thermo Fisher Scientific (1 mentions) Sdf 1α, by R&D Systems (1 mentions) Sebacic acid, by Merck Group (1 mentions) Glycerol, by Thermo Fisher Scientific (1 mentions) 1 1 1 3 3 3 hexafluoro 2 propanol hfip, by Merck Group (1 mentions)

6 protocols using recombinant human sdf 1α

1

Investigating IGF1R and CXCR4 Signaling

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The immunoprecipitation experimental procedures were as previously described54 (link). Cultures were treated with recombinant human IGF1 (Invitrogen) at 300 ng/mL80 (link) and/or recombinant human SDF-1α (R&D System) at 100 nM81 (link). First, hDSCs were collected and lysed in lysis buffer (50 mM Tris-HCl, pH 7.5, 1% NP-40, 150 mM NaCl, 0.5% sodium deoxycholate, and protease inhibitor). The cell lysate (300 μg) was incubated with protein A/G-agarose beads at 4 °C for 6 hours. Then, antibodies against IGF1R (Santa Cruz Biotechnology), CXCR4 (R&D System), phosphotyrosine (pY) (p-Tyr-100, Cell Signaling), Giα2 (T-19, Santa Cruz Biotechnology) and Gβ (M-14, Santa Cruz Biotechnology), or control antibody IgG (anti-hemagglutinin clone F-7, Santa Cruz Biotechnology) were added and reacted for 6 hours at 4 °C. These immunocomplexes were incubated on protein A/G-agarose beads at 4 °C overnight. After three washes with lysis buffer, the immunocomplexes were then examined by western blot with anti-IGF1R, anti-phosphotyrosine, anti-CXCR4 antibodies, anti- Giα2 and Gβ, or anti-control IgG.
Lee H.T., Chang H.T., Lee S., Lin C.H., Fan J.R., Lin S.Z., Hsu C.Y., Hsieh C.H, & Shyu W.C. (2016). Role of IGF1R+ MSCs in modulating neuroplasticity via CXCR4 cross-interaction. Scientific Reports, 6, 32595.
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2

Immunophenotyping of Hematopoietic Stem Cells

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PE-conjugated rat anti-human CD184/CXCR4 (clone 1D9, isotype control rat IgG2a,κ), FITC-conjugated mouse anti-Rac1 (clone 102/Rac1, isotype control mouse IgG2,b), APC-conjugated mouse anti-human CD34 (clone 581, isotype mouse IgG1,κ), PE-conjugated mouse anti-human CD38 (clone HIT2, isotype control mouse IgG1,κ) and APC-conjugated mouse anti-human CD45 (clone Hl30, isotype mouse IgG1,κ) were purchased from BD Biosciences (San Diego, CA, USA). Blocking reagents human gamma globulin and mouse gamma globulin were purchased from Jackson ImmunoResearch Laboratories Incorporated (West Grove, PA, USA). BD Cytofix™ fixation buffer was purchased from BD Biosciences. Recombinant human SDF-1α was purchased from R&D Systems. FITC-conjugated Cholera toxin B subunit (CTxB) and methyl-β-cyclodextrin (MβCD) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Rac1 inhibitor NSC23766 was purchased from BioVision (Milpitas, CA, USA).
Capitano M.L., Hangoc G., Cooper S, & Broxmeyer E. (2015). Mild heat treatment primes human CD34+ cord blood cells for migration towards SDF-1α and enhances engraftment in an NSG mouse model. Stem cells (Dayton, Ohio), 33(6), 1975-1984.
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3

Multicolor Fluorescence Labeling Assay

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Fluo-4 AM, Rhod-2 AM, MitoTracker Red CM-H2Xros, and carboxyfluorescein succinimidyl ester (CFSE) were purchased from Molecular Probes (Thermo Fisher Scientific). Recombinant human SDF-1α, NF340 (P2Y11 antagonist), NF546 (P2Y11 agonist), 5-BDBD (P2X4 antagonist), CSC (A2a antagonist), MRS1754 (A2b antagonist), and H89 (PKA antagonist) were from R&D Systems, and cAMP-AM was from BIOLOG Life Science Institute. In all experiments, 5-BDBD, NF340, and NF546 were used at concentrations (1-10 μM) at which they were reported to be specific for the respective receptor (36 (link), 64 (link), 65 (link)). Anti-human CD4-allophycocyanin (clone: OKT4) and anti-human CD69 FITC (clone: FN50) antibodies were purchased from Biolegend. Mouse anti-human CD3 (clone: HIT3a) antibodies were from BD Pharmingen. All other reagents were purchased from Sigma-Aldrich unless stated otherwise.
Ledderose C., Bromberger S., Slubowski C.J., Sueyoshi K., Aytan D., Shen Y, & Junger W.G. (2020). The purinergic receptor P2Y11 choreographs the polarization, mitochondrial metabolism, and migration of T lymphocytes. Science signaling, 13(651), eaba3300.
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4

Gelatin Sponge Delivery of SDF-1α

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Commercially available gelatin sponge (Spongostan®; Ferrosan Medical Devices, Denmark) was resized to the defect size (2 × 2 × 2 mm3, W × L × D) and loaded via absorption with 20 μl PBS solution containing three different doses of recombinant human SDF‐1α (R&D systems, Abingdon, UK): 100, 200 and 400 ng SDF‐1α (Cao et al., 2013; Du et al., 2012). All procedure was performed under aseptic conditions.
Cai X., Yang F., Walboomers X.F., Wang Y., Jansen J.A., van den Beucken J.J, & Plachokova A.S. (2018). Periodontal regeneration via chemoattractive constructs. Journal of Clinical Periodontology, 45(7), 851-860.
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5

Synthesis and Characterization of PGS Scaffold

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Poly(glycerol‐sebacate) (PGS) prepolymer was synthesized with sebacic acid purchased from Sigma‐Aldrich (St. Louis, MO) and glycerol purchased from Fisher Scientific (Waltham, MA) using a published protocol.23 Type I collagen was purchased commercially from Elastin Products Company, Inc (Owensville, MO). Silk fibroin was extracted from raw silk purchased from Haian Silk Company (Nantong, China) according to published methods.24 Human bone marrow derived CD34+ cells were purchased from ATCC (Manassas, VA). 1,1,1,3,3,3‐hexafluoro‐2‐propanol (HFIP) and fibronectin were purchased from Sigma‐Aldrich (St. Louis, MO). Recombinant human SDF‐1α, human sydecan‐4 and Quanti SDF‐1α ELISA kit were obtained from R&D Systems (Minneapolis, MN). EGM™‐2 Endothelial Cell Growth Medium‐2 BulletKit™ was purchased from Lonza (Walkersville, MD). Quant‐iT™ PicoGreen™ dsDNA Assay Kit, 4′,6‐diamidino‐2‐phenylindole (DAPI), Rhodamine Phalloidin and SYTOX™ Green Nucleic Acid Stain were purchased from Invitrogen (Waltham, MA).
Wu Y., Yazdani S.K., Bolander J.E, & Wagner W.D. (2022). Syndecan‐4 and stromal cell‐derived factor‐1 alpha functionalized endovascular scaffold facilitates adhesion, spreading and differentiation of endothelial colony forming cells and functions under flow and shear stress conditions. Journal of Biomedical Materials Research. Part B, Applied Biomaterials, 111(3), 538-550.
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6

Quantifying EPC Adhesion to SDF-1α-Stimulated EC

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1 x 105 cells/well of HUVEC were cultured in the 24-well glass plates pre-coated with 1% gelatin and cells reached 100% confluence one-day later. The HUVEC monolayers were stimulated with recombinant human SDF-1α (R & D Systems, Minneapolis, MN) or BSA at 100 ng/mL for 8-hour. Nanocarriers-coated DsRed+ EPC (1 x 105) were suspended in 1 mL basal EGM2 medium. To test the effect of E-selectin antagonist on EPC adhesion to SDF-1α-stimulated EC, E-selectin neutralizing antibody (BBA1; R & D Systems) or isotype-matched control antibody (2 μg/mL) were added into EC monolayer and incubated for 15-min before adding nanocarriers-coated EPC. HUVEC-EPC were co-cultured for 1-hour at 37°C. Unbounded EPC were washed out twice with 1 mL PBS. Fluorescence signals derived from adherent DsRed+ EPC were measured and quantified by fluorescence scanner (GE Typhoon Trio, Piscataway, NJ).
Liu Z.J., Daftarian P., Kovalski L., Wang B., Tian R., Castilla D.M., Dikici E., Perez V.L., Deo S., Daunert S, & Velazquez O.C. (2016). Directing and Potentiating Stem Cell-Mediated Angiogenesis and Tissue Repair by Cell Surface E-Selectin Coating. PLoS ONE, 11(4), e0154053.
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