Mouse GRO-alpha (CXCL1)Recombinant

////Mouse GRO-alpha (CXCL1)Recombinant

Mouse GRO-alpha (CXCL1)Recombinant

$70.00$2,700.00

SKU: RKP12850 Category: Tags: , , , , , , , ,

accession P12850


Source Optimized DNA sequence encoding mouse GRO alpha mature chain was expressed in Escherichia Coli.
Molecular weight Native mouse CXCL1/GRO-alpha is generated by the proteolytic removal of the signal peptide and propeptide. The molecule has a calculated molecular mass of approximately 8 kDa. Recombinant mouse CXCL1/GRO-a is a monomer protein consisting of 72 amino acid residue subunits, and migrates as an approximately 8 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 MIPATRSLLC AALLLLATSR LATGAPIANE LRCQCLQTMA GIHLKNIQSL KVLPSGPHCT QTEVIATLKN GREACLDPEA PLVQKIVQKM LKGVPK
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 mouse GRO-alpha/CXCL1was 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.


Biological Process Inflammatory-response
Molecular function Cytokine
Molecular function Growth-factor

Methods

Leukocyte adhesion in CXCL1-stimulated cremaster muscle venules.

recombinant murine CXCR2 chemokine CXCL1
  • The number of adherent cells (mean ± SEM) was assessed in exteriorized cremaster muscle venules of RAGE-/- mice (10 venules in 6 mice), Icam1mice (11 venules in 5 mice), untreated (12 venules in 7 mice), anti-LFA-1 antibody (10 venules in 4 mice) or isotype antibody (9 venules in 3 mice) treated wild type mice before and five minutes after systemic injection of CXCL1 (keratinocyte-derived chemokine, KC; 600 ng/mouse).

Leukocyte adhesion (number of adherent cells/mm2) in mouse cremaster muscle venules and leukocyte transmigration (perivascular neutrophils/mm2 surface area) in cremaster muscle whole mounts in response to glucose during trauma-induced inflammation.

recombinant murine chemokine CXCL1
  • To dissect the role of the chemokine pathway in this model, WT mice were either pretreated with 4 μg PTx 3 h prior to cremaster exteriorization and glucose stimulation or coinjected with 600 ng CXCL1 and glucose and compared to injection of normal saline.

Flow Cytometric Analysis

  • For flow cytometric analysis of surface markers and intracellular cytokines, enriched C4+ T cells, and single-cell suspensions of lungs were stained with optimal concentrations of the following specific antibodies against surface molecules and intracellular cytokines: C8 α-V450, C4-V500, C11c-FITC, C62L-APC, Gr-1-APC, IFNγ-APC from , , , , and MHCII(I-A/I-E)-eFluor780 from eBioscience, C44-FITC, and IFNγ-PE fromand IL-22-PE from& .
  • Fluorescence intensity was measured by using a FacsCantoII® flow cytometer equipped with a 405 nm, 488 nm and 633 nm laser.
  • Analysis was performed utilizing the FCSExpress4 program (DeNovo™ Software) gated on leukocytes identified by the forward-scatter/side-scatter profile and further characterization as specified in the figures.

In vivo experiments

  • For adoptive transfer studies, classical monocytes were isolated from BM by FACS sorting using antibodies to CD45, CD115, and Gr1.
  • After labelling the monocytes with CFSE, 106 cells were adoptively transferred by tail vein injection to .
  • Twenty-four hours after transfer, aortas and hearts of recipient mice were collected for further analysis.
  • For monocyte mobilization studies, rmCXCL1/KC was injected i.v.
  • at 40 µg/kg.
  • After 1 h, blood was drawn and mice were sacrificed to harvest bones and spleens.
  • Serum CXCL1 in mice was neutralized by daily administration of 5 µg of anti-CXCL1 antibody (clone 124014) or IgG isotype control (clone 54447, both& ) for 1 week and for 3 consecutive weeks every other day.

Elimination of 12/15-LO activity alters chemokine homeostasis.

recombinant CXCL1
  • CXCL1 and CXCL2 concentrations in the plasma was measured in WT mice (black bars) and Alox15mice (white bars) under baseline conditions and 3 h after LPS inhalation (n = 3 to 4).

Sdc-3 deletion reduces neutrophil recruitment in CXCL1-injected joints.

recombinant murine CXCL1
  • Sdc-3 null and wild-type mice were injected intra-articularly with 3 μg per knee joint of recombinant murine CXCL1 or PBS and after 4 hours processed for histology.