Mice were given antibiotics in drinking water as described previously10 (link), 15 (link). Mice were given broad-spectrum antibiotics (ampicillin 1 g/L, neomycin sulfate 1 g/L, metronidazole 1 g/L, and vancomycin 0.5 g/L) in drinking water for 10–14 days. Antibiotic therapy was stopped 3 days prior to infection. PRR ligands were prepared as described previously and administered by oral gavage10 (link). Anti-GM-CSF (BioLegend), anti-CXCL2 (R&D Systems), anti-CXCL1 (R&D Systems), and isotype control, rGM-CSF (BioLegend) or rIL-17A (Thermo Fisher Scientific), were administered intranasally concomitant with S. pneumoniae or K. pneumoniae inoculation, unless stated otherwise. Anti-IL-17A (R&D Systems) was administered 3 days prior to infection and concomitant with infection via the intraperitoneal route.
Anti cxcl2
Manufactured by R&D Systems
Anti-CXCL2 is a recombinant antibody that binds to and neutralizes the CXCL2 chemokine. CXCL2 is a chemotactic factor that is involved in the recruitment and activation of neutrophils. This antibody can be used to study the role of CXCL2 in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Anti cxcl1, by R&D Systems (2 mentions)
Anti il 17a, by R&D Systems (1 mentions)
Rgmcsf, by BioLegend (1 mentions)
Anti gm csf, by BioLegend (1 mentions)
Tumor necrosis factor (tnf), by R&D Systems (1 mentions)
Anti cxcl5, by R&D Systems (1 mentions)
Il 1β, by R&D Systems (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
Matrigel, by BD (1 mentions)
Anti ccl5, by R&D Systems (1 mentions)
4 protocols using anti cxcl2
1
Antibiotic Modulation of Respiratory Infection
2
Neutrophil Extravasation Regulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were anaesthetized with 3% isoflurane and injected i.s. with 300 ng TNF or 50 ng IL-1β (both R&D Systems), whereas control mice received 400 μl PBS (2-4 hr incubation). For the analysis of total neutrophil extravasation blocking anti-CXCL1, anti-CXCL2, anti-CXCL5 (all R&D Systems) or anti-MIF mAbs (kindly provided by Dr Christian Weber, Ludwig Maximilians University of Munich, Germany) or corresponding isotype control mAbs (30 μg/mouse, R&D Systems) were injected i.s. together with TNF. For IVM analyses mAbs were applied as described in the corresponding section below.
3
Neutrophil and Monocyte Recruitment in Peritonitis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Peritonitis was induced by intraperitoneal (i.p.) injections of 300 ng TNF in WT mice or Cxcl2−/−, Ackr1−/− or corresponding control chimeras, whereas non-inflamed control mice received 1 mL of PBS. Blocking anti-CXCL1, anti-CXCL2, anti-CXCR2 mAbs (R&D Systems) or corresponding isotype control mAbs (3 mg/kg, R&D Systems) were administered i.v. 10 min prior to TNF. 4 hr after TNF or PBS administration, mice were culled and subjected to peritoneal lavages using 5 mL PBS containing 5 mM EDTA (Sigma-Aldrich) and 0.25% BSA. The peritoneal exudates were stained for the leukocyte marker CD45, the neutrophil marker Ly6G (both Biolegend) and the monocyte and macrophage marker CD115 (Thermo Fisher Scientific) and total numbers of infiltrating neutrophils and monocytes per peritoneal cavity were determined by flow cytometry.
4
Transwell Assay for FLS Migration and Invasion
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the transwell migration and invasion assay, FLS from WT (Epas1+/+) or Epas1+/− mice (C57BL/6) were seeded on membranes of inserts with 8.0 μm pores (Corning Costar, Corning, NY, USA ) and the lower chambers filled with CM, which served as the chemoattractant. Serum-free CM was prepared from primary cultures of chondrocytes or FLS isolated from WT or Epas1+/− mice (C57BL/6). For neutralization of chemokines, 0.5 μg anti-IgG (control), anti-CXCL2, or anti-CCL5 (R&D Systems) were added to the lower chamber. For the invasion assay, inserts of the transwell were precoated with matrigel (BD Bioscience, San Jose, CA, USA). After 24 hours of incubation, transwell inserts were fixed with 4 % paraformaldehyde for 10 minutes and stained with 0.05 % crystal violet. Cells on the bottom layer were captured in five randomly selected fields at 200× magnification using a light microscope and quantified with Image J software (NIH, shareware) .
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!