Human GRO-alpha (CXCL1)Recombinant

/, CXC chemokines/Human GRO-alpha (CXCL1)Recombinant

Human GRO-alpha (CXCL1)Recombinant

$70.00$2,700.00


accession P09341


Source Optimized DNA sequence encoding HumanGRO alphamature chain was expressed in Escherichia Coli.
Molecular weight Nativehuman GRO-alpha isgenerated by the proteolytic removal of the signal peptideand propeptide. The molecule has a calculated molecular mass of approximately8 kDa. Recombinant GRO-a is a disulfide-linked homodimeric protein consisting of amino acid residue subunits, andmigrates 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 theability to chemoattract human neutrophils using a concentration range of.0-50.0 ng/ml.

Protein Sequence MARAALSAAP SNPRLLRVAL LLLLLVAAGR RAAGASVATE LRCQCLQTLQ GIHPKNIQSV NVKSPGPHCA QTEVIATLKN GRKACLNPAS PIVKKIIEKM LNSDKSN
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 GRO-alpha/CXCL1was lyophilized from a.2μm filtered concentrated (1mg/ml) solution inmM NaCl,mM PB, pH.0.
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 P25025
Interactor Q16570
Interactor P25024
Biological Process Inflammatory-response
Molecular function Cytokine
Molecular function Growth-factor

Methods

Immunofluorescence Confocal microscopy

  • For co-localization studies examining Duffy with early endosomal antigen (EEA) and lysosomal associated membrane protein (LAMP), DIH cells were grown in 24 well plates containing 12 mm glass coverslips.
  • Cells were stimulated with or without 100 ng/mL human recombinant CXCL1/GRO-α or 50 ng/ml PDGF-BB for various time points, and then fixed with 2% paraformaldehyde in PBS for 1 hr and permeabilized with 0.1% Triton in PBS for 30 min.
  • Cells were washed in 0.5% BSA in PBS , blocked with 2% BSA in PBS for 30 min, and incubated with primary antibodies mouse anti-Duffy 2C3 Fy6, rabbit anti-EEA (1∶100 Abcam, , ) and rabbit anti-LAMP (1∶100 Abcam, , ) for 1 hr at RT.
  • Cells were washed 4 x with PBB then were stained with goat anti-mouse Cy3 (1∶1000 ), goat anti-rabbit Alexa 488 (1∶500) and phalloidin-Alexa 647 (1∶250) for 1 hr at RT.
  • Cells were processed and imaged for confocal microscopy…
  • For co-localization studies examining Duffy with early endosomal antigen (EEA) and lysosomal associated membrane protein (LAMP), DIH cells were grown in 24 well plates containing 12 mm glass coverslips.
  • Cells were stimulated with or without 100 ng/mL human recombinant CXCL1/GRO-α or 50 ng/ml PDGF-BB for various time points, and then fixed with 2% paraformaldehyde in PBS for 1 hr and permeabilized with 0.1% Triton in PBS for 30 min.
  • Cells were washed in 0.5% BSA in PBS , blocked with 2% BSA in PBS for 30 min, and incubated with primary antibodies mouse anti-Duffy 2C3 Fy6, rabbit anti-EEA (1∶100 Abcam, , ) and rabbit anti-LAMP (1∶100 Abcam, , ) for 1 hr at RT.
  • Cells were washed 4 x with PBB then were stained with goat anti-mouse Cy3 (1∶1000 ), goat anti-rabbit Alexa 488 (1∶500) and phalloidin-Alexa 647 (1∶250) for 1 hr at RT.
  • Cells were processed and imaged for confocal microscopy as described above.

Read more