Human SDF-1 alpha Recombinant

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Human SDF-1 alpha Recombinant

$70.00$3,500.00


accession P48061


Source Optimized DNA sequence encoding Human Stromal Cell-Derived Factor-1 alpha mature chain was expressed in Escherichia Coli.
Molecular weight Native human SDF-1 alpha, generated by the proteolytic removal of the signal peptide and propeptide,the molecule has a calculated molecular mass of approximately8 kDa. Recombinant SDF-1 alpha is a disulfide-linked monomer protein consisting of69 amino acid residue subunits, migrates as an approximately8 kDa protein under non-reducing and reducing conditions in SDS-PAGE.
Purity >97%, as determined by SDS-PAGE and HPLC
Biological Activity Determined by its ability to chemoattract human U937 expressing CXCR4 Determined by calcium flux with human U937 cells.
Protein Sequence MNAKVVVVLV LVLTALCLSD GKPVSLSYRC PCRFFESHVA RANVKHLKIL NTPNCALQIV ARLKNNNRQV CIDPKLKWIQ EYLEKALNK R FKM
Endotoxin Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than.1 ng/µg(1EU/µg).
Presentation Recombinant SDF-1 alpha was lyophilized from a.2μm filtered PBS, pH.4.
Reconstitution A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than.1 mg/mL. This solution can then be diluted into other buffers.
Storage The lyophilized protein is stable for at least years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at° -° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.


Interactor P15172
Biological Process Chemotaxis
Molecular function Cytokine
Molecular function Growth-factor

Methods

Migration assay

  • Peripheral blood samples at TP2 in EDTA tubes are diluted with PBS supplemented with 2 mM EDTA and 0.5% BSA (v/v, 1∶1); 30 mL of diluted blood solution is then loaded in a 50-mL LeucoSep tubes with 15 mL of Histopaque 1077 and centrifuged at 850×g for 25 min.
  • The buffy coat layers containing white blood cells are collected and washed twice with sterile PBS.
  • Cells are then resuspended in EBM-2 supplemented with 0.1% BSA.
  • 0.5–1×107 cells/mL of cells are loaded onto the upper part of ThinCert™ tissue culture polystyrene inserts (452.4 mm2 culture surface, 3.0 µm pore size , , ) which are pre-assembled on 6-well plates containing medium as above with or without 100 ng/mL of recombinant human SDF-1α or β-NGF , respectively.
  • Cells are then incubated at 37°C in a humidified incubator with 5% CO2 for 18 hrs.
  • Cells from both parts of the transwell inserts are dissociated…
  • Peripheral blood samples at TP2 in EDTA tubes are diluted with PBS supplemented with 2 mM EDTA and 0.5% BSA (v/v, 1∶1); 30 mL of diluted blood solution is then loaded in a 50-mL LeucoSep tubes with 15 mL of Histopaque 1077 and centrifuged at 850×g for 25 min.
  • The buffy coat layers containing white blood cells are collected and washed twice with sterile PBS.
  • Cells are then resuspended in EBM-2 supplemented with 0.1% BSA.
  • 0.5–1×107 cells/mL of cells are loaded onto the upper part of ThinCert™ tissue culture polystyrene inserts (452.4 mm2 culture surface, 3.0 µm pore size , , ) which are pre-assembled on 6-well plates containing medium as above with or without 100 ng/mL of recombinant human SDF-1α or β-NGF , respectively.
  • Cells are then incubated at 37°C in a humidified incubator with 5% CO2 for 18 hrs.
  • Cells from both parts of the transwell inserts are dissociated with Accutase and collected into FACS tubes.
  • Following washing using PBS, migrated and non-migrated cells are stained with KDR-FITC, CD133/2 (293C3)-APC, CD34 (8G12)-PE-Cy7 and CXCR4 (CD184)-PE antibodies for flow cytometry analysis as described above.

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Migration assay

  • Peripheral blood samples at TP2 in EDTA tubes are diluted with PBS supplemented with 2 mM EDTA and 0.5% BSA (v/v, 1∶1); 30 mL of diluted blood solution is then loaded in a 50-mL LeucoSep tubes with 15 mL of Histopaque 1077 and centrifuged at 850×g for 25 min.
  • The buffy coat layers containing white blood cells are collected and washed twice with sterile PBS.
  • Cells are then resuspended in EBM-2 supplemented with 0.1% BSA.
  • 0.5–1×107 cells/mL of cells are loaded onto the upper part of ThinCert™ tissue culture polystyrene inserts (452.4 mm2 culture surface, 3.0 µm pore size , , ) which are pre-assembled on 6-well plates containing medium as above with or without 100 ng/mL of recombinant human SDF-1α or β-NGF , respectively.
  • Cells are then incubated at 37°C in a humidified incubator with 5% CO2 for 18 hrs.
  • Cells from both parts of the transwell inserts are dissociated…
  • Peripheral blood samples at TP2 in EDTA tubes are diluted with PBS supplemented with 2 mM EDTA and 0.5% BSA (v/v, 1∶1); 30 mL of diluted blood solution is then loaded in a 50-mL LeucoSep tubes with 15 mL of Histopaque 1077 and centrifuged at 850×g for 25 min.
  • The buffy coat layers containing white blood cells are collected and washed twice with sterile PBS.
  • Cells are then resuspended in EBM-2 supplemented with 0.1% BSA.
  • 0.5–1×107 cells/mL of cells are loaded onto the upper part of ThinCert™ tissue culture polystyrene inserts (452.4 mm2 culture surface, 3.0 µm pore size , , ) which are pre-assembled on 6-well plates containing medium as above with or without 100 ng/mL of recombinant human SDF-1α or β-NGF , respectively.
  • Cells are then incubated at 37°C in a humidified incubator with 5% CO2 for 18 hrs.
  • Cells from both parts of the transwell inserts are dissociated with Accutase and collected into FACS tubes.
  • Following washing using PBS, migrated and non-migrated cells are stained with KDR-FITC, CD133/2 (293C3)-APC, CD34 (8G12)-PE-Cy7 and CXCR4 (CD184)-PE antibodies for flow cytometry analysis as described above.

Read more

Migration assay

  • Peripheral blood samples at TP2 in EDTA tubes are diluted with PBS supplemented with 2 mM EDTA and 0.5% BSA (v/v, 1∶1); 30 mL of diluted blood solution is then loaded in a 50-mL LeucoSep tubes with 15 mL of Histopaque 1077 and centrifuged at 850×g for 25 min.
  • The buffy coat layers containing white blood cells are collected and washed twice with sterile PBS.
  • Cells are then resuspended in EBM-2 supplemented with 0.1% BSA.
  • 0.5–1×107 cells/mL of cells are loaded onto the upper part of ThinCert™ tissue culture polystyrene inserts (452.4 mm2 culture surface, 3.0 µm pore size , , ) which are pre-assembled on 6-well plates containing medium as above with or without 100 ng/mL of recombinant human SDF-1α or β-NGF , respectively.
  • Cells are then incubated at 37°C in a humidified incubator with 5% CO2 for 18 hrs.
  • Cells from both parts of the transwell inserts are dissociated…
  • Peripheral blood samples at TP2 in EDTA tubes are diluted with PBS supplemented with 2 mM EDTA and 0.5% BSA (v/v, 1∶1); 30 mL of diluted blood solution is then loaded in a 50-mL LeucoSep tubes with 15 mL of Histopaque 1077 and centrifuged at 850×g for 25 min.
  • The buffy coat layers containing white blood cells are collected and washed twice with sterile PBS.
  • Cells are then resuspended in EBM-2 supplemented with 0.1% BSA.
  • 0.5–1×107 cells/mL of cells are loaded onto the upper part of ThinCert™ tissue culture polystyrene inserts (452.4 mm2 culture surface, 3.0 µm pore size , , ) which are pre-assembled on 6-well plates containing medium as above with or without 100 ng/mL of recombinant human SDF-1α or β-NGF , respectively.
  • Cells are then incubated at 37°C in a humidified incubator with 5% CO2 for 18 hrs.
  • Cells from both parts of the transwell inserts are dissociated with Accutase and collected into FACS tubes.
  • Following washing using PBS, migrated and non-migrated cells are stained with KDR-FITC, CD133/2 (293C3)-APC, CD34 (8G12)-PE-Cy7 and CXCR4 (CD184)-PE antibodies for flow cytometry analysis as described above.

Read more

Receptors involved in the crosstalk between CXCL12 and HER1.

200 ng/mL CXCL12
  • 5637 or HeLa cells were cultured alone, or in the presence of 200 ng/mL CXCL12 or 25 ng/mL HB-EGF.

TOSE cells are hypermethylated at the CXCL12 promoter (A) CXCL12 RNA levels were measured by quantitative RT-PCR.

100ng/ml CXCL12
  • CXCL12 protein levels measured by ELISA (inset).