After blocking (×1 PBS, 2 % normal goat serum, and 0.1 % Triton X-100) for 1 h, the sections were incubated overnight at 4 °C with a mixture of any two of the following primary antibodies: anti-CXCL5 (1:50; abcam), anti-ED1 (1:100; Chemicon, Temecula, CA), anti-O4 IgM (1:100; Chemicon), anti-GFAP (1:200; Chemicon), anti-neuronal nuclear antigen (NeuN) (1:200; Millipore Bioscience Research Reagents), and anti-RECA (1:100; abcam). The sections were washed three times with 0.1 M PBS and incubated with Alexa Fluor 594 anti-mouse IgG/IgM or Alexa Fluor 488 anti-rabbit IgG (1:400; Invitrogen) for 1 h at room temperature. The fluorescence signals were detected, and the results were recorded using a microscope (E400; Nikon Instech) at excitation–emission wavelengths of 596–615 nm (Alexa Fluor 594, red) and 470–505 nm (Alexa Fluor 488, green) [18 (link), 23 (link)].
Anti cxcl5
Manufactured by Abcam
Sourced in United States
Anti-CXCL5 is a laboratory reagent used for the detection and quantification of the CXCL5 protein. CXCL5 is a chemokine involved in the immune response. This antibody can be used in various immunoassay techniques to measure CXCL5 levels in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti gfap, by Merck Group (1 mentions)
Anti reca, by Abcam (1 mentions)
Alexa fluor 488 anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Anti cxcr2, by Abcam (1 mentions)
Anti cd206, by Abcam (1 mentions)
Anti cd8, by Abcam (1 mentions)
Opal 7 colour manual ihc kit, by PerkinElmer (1 mentions)
Vectra polaris automated quantitative pathology imaging system, by Akoya Biosciences (1 mentions)
Anti cxcl8, by Abcam (1 mentions)
Anti cxcl1, by Abcam (1 mentions)
Anti cd68, by Abcam (1 mentions)
Anti rad51, by Abcam (1 mentions)
Anti phospho histone h2a x ser139, by Merck Group (1 mentions)
Anti g0s2, by Proteintech (1 mentions)
Anti 53bp1, by Abcam (1 mentions)
Anti rnf168, by Abcam (1 mentions)
Anti p65, by Cell Signaling Technology (1 mentions)
Anti mmp9, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
4 protocols using anti cxcl5
1
Immunofluorescence Labeling of Neural Markers
2
Multiplex Immunohistochemistry of Pancreatic Cancer
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Fluorescent mIHC was performed in serial sections of FFPE tumor tissue from each PDAC patient using the Opal 7-Colour Manual IHC Kit (PerkinElmer Hopkinton, Massachusetts, USA) according to the manufacturer’s protocol and as described in our previous article. In short, sections experienced primary antibodies including Rabbit monoclonal antibodies anti-CD68 (1:500, Abcam, Cambridge, MA, USA), anti-CXCR2 (1:500, Abcam), anti-CD206 (1000, Abcam), anti-CXCL1 (1:200, Abcam), anti-CXCL5 (1:200, Abcam), anti-CXCL8 (1:500, Abcam) and anti-CD8 (1:2000, Abcam), followed by HRP-conjugated secondary antibody and fluorescent dyes (Opal520, Opal570, Opal620, Opal690, and DAPI). Stained sections were scanned and processed by a Vectra PolarisTM Automated Quantitative Pathology Imaging System (AKOYA Biosciences, PerkinElmer, Massachusetts, USA).
3
Antibody Immunostaining Protocol
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The following antibodies were used in this study: an anti-G0S2 (dilution 1:200; Proteintech, IL, USA); an anti-phospho-Histone H2A.X (Ser139) (dilution1:1000; EMD Millipore, Billerica, MA, USA); anti-53BP1 (dilution1:500), anti-RNF168 (dilution1:500), anti-CXCL5 (dilution 1:1000) and anti-Rad51 (dilution 1:1000)(Abcam, Cambridge, MA, USA). The secondary antibodies were from Jackson ImmunoResearch Laboratories (West Grove, PA, USA). Cell culture media and other reagents were from Invitrogen (Carlsbad, CA, USA), Sigma-Aldrich (St. Louis, MO, USA) or Peprotech (Rocky Hill, NJ, USA).
4
Western Blot Analysis of Inflammatory Markers
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Cells and tissue samples were lysed with radioimmunoprecipitation assay (RIPA) buffer containing a protease inhibitor cocktail (B14001, Bimake, Houston, TX, USA). Protein concentrations were measured by bicinchoninic acid assay. Equal amounts of protein mixtures (30 µg) were run on sodium dodecyl sulfate-PAGE, and the proteins were transferred to polyvinylidene fluoride (PVDF) membranes (Millipore, Billerica, MA, USA) and incubated in PBS-Tween 20 with 5% nonfat milk for 1 hour at room temperature. The blot was incubated with primary antibodies, including anti-CXCL5 (1:100, no. ab198505, Abcam), anti-MMP9 (1:1000, no. 3852, Cell Signaling Technology, Danvers, MA, USA), anti-p65 (1:1000, no. 8242, Cell Signaling Technology) and anti-β-actin (1:20,000, no. KC-5A08, Shanghai Kangcheng, Shanghai, China) overnight at 4°C. Secondary antibodies conjugated to horseradish peroxidase were incubated with the blot for 1 hour at room temperature. The proteins revealed by western blotting were visualized by using enhanced chemiluminescence reagents (Servicebio, Wuhan, China). The densities of bands were analyzed by ImageJ software (National Institutes of Health, Bethesda, MD) and normalized to expression of GAPDH.
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