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Af3059

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AF3059 is a laboratory equipment product manufactured by R&D Systems. It is designed for general laboratory use.

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Lab products found in correlation

Af2065, by R&D Systems (3 mentions) Alexafluor 488 chicken anti goat igg, by Thermo Fisher Scientific (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Alexa fluor 488 goat anti rat igg, by Thermo Fisher Scientific (1 mentions) Proteinase k, by Agilent Technologies (1 mentions) Intracellular staining kit, by Thermo Fisher Scientific (1 mentions) Anti mouse cd16 cd32 clone 93, by Thermo Fisher Scientific (1 mentions) Accuri c6, by BD (1 mentions) Ab52642, by Abcam (1 mentions) Ab9234, by Merck Group (1 mentions) Calnexin h 70, by Santa Cruz Biotechnology (1 mentions) Ab31721, by Abcam (1 mentions) Ab15106, by Abcam (1 mentions) Pvdf transfer membrane, by Thermo Fisher Scientific (1 mentions) Cl xposure film, by Thermo Fisher Scientific (1 mentions) Ab6640, by Abcam (1 mentions) Pierce ecl substrate, by Thermo Fisher Scientific (1 mentions) Dm6 b, by Leica (1 mentions)

5 protocols using af3059

1

Immunohistochemical Analysis of Corneal Inflammation

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Whole eyes were fixed in 10% phosphate buffered formalin, paraffin embedded and sectioned. PASH and GMS staining were performed by the CWRU Visual Science Research Center histology core. For immunohistochemistry, sections were treated with proteinase K (Dako 2015-011, Carpinteria, CA) and blocked in 1.5% serum. Mouse S100A9 polyclonal Ab (2 µg/ml, R&D Systems AF2065, Minneapolis, MN), Mouse S100A8 polyclonal Ab (2 µg/ml R&D Systems AF3059), or Anti-mouse neutrophil antibody NIMP-R14 (20 µg/ml) were used, followed by staining with Alexafluor 488 Chicken anti-goat IgG (1:2000, Life Technologies A2467, Grand Island, NY) or Alexafluor 488 Goat anti-rat IgG (1:250, Life Technologies A11006). Slides were imaged at 200–400X. Neutrophil quantification from histological sections was obtained by measuring % NIMP-R14 positive area/cornea using Metamorph software.
Clark H.L., Jhingran A., Sun Y., Vareechon C., Carrion S.D., Skaar E.P., Chazin W.J., Calera J.A., Hohl T.M, & Pearlman E. (2015). Zinc and Manganese Chelation by Neutrophil S100A8/A9 (Calprotectin) Limits Extracellular Aspergillus fumigatus Hyphal Growth and Corneal Infection. Journal of immunology (Baltimore, Md. : 1950), 196(1), 336-344.
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2

Neutrophil Characterization in Cornea

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Corneas were dissected at 24–48h pi and treated with 1X collagenase for 1–2 h. Cell suspensions were washed in 1X PBS + 5% FBS, incubated with anti-mouse CD16/CD32 (clone 93, eBioscience 16–0161-86, San Diego, CA)(to block Fc receptors) and stained with anti-mouse neutrophil antibody NIMP-R14-PE (abcam ab125259, Cambridge, MA) or isotype for 1h and live/dead fixable far read stain (Thermo Fisher). For intracellular staining, cells were fixed and permeabilized using an intracellular staining kit (eBioscience). Cells were incubated with Mouse S100A9 polyclonal Ab ( R&D Systems AF2065), Mouse S100A8 polyclonal Ab ( R&D Systems AF3059), or isotype for 45 min, washed and incubated with Alexafluor 488 Chicken anti-goat IgG (1:2000, Life Technologies A2467) for 30 min, washed, resuspended in FACS buffer and analyzed in a BD Accuri C6 (San Jose, CA). Analysis was performed using Accuri C6 software. Cells were gated on FSC/SSC, followed by live cells.
Clark H.L., Jhingran A., Sun Y., Vareechon C., Carrion S.D., Skaar E.P., Chazin W.J., Calera J.A., Hohl T.M, & Pearlman E. (2015). Zinc and Manganese Chelation by Neutrophil S100A8/A9 (Calprotectin) Limits Extracellular Aspergillus fumigatus Hyphal Growth and Corneal Infection. Journal of immunology (Baltimore, Md. : 1950), 196(1), 336-344.
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3

Comprehensive Alzheimer's Protein Analysis

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The following primary antibodies were used: anti-Aβ mouse monoclonal antibody 6E10 (MAB5206; Chemicon), anti-mouse S100A9 and S100A8 (AF2065, AF3059; R&D systems), S100B (ab52642;Abcam), GAPDH (Abfrontier), anti-Amyloid Oligomer, Aβ, (AB9234; Millipore),p-Tau (Ser404) (sc-12952;Santa Cruz Biotech.), Phospho-PHF-tau (S202/T205, AT8) (NM1020;Pierce), Anti-PhosphoTau (S396;PHF-13) (ab24716;Abcam), Tau (C-17) (sc-1995;Santa Cruz Biotechnology), Calnexin (H-70) (sc-11397;Santa Cruz Biotechnology), and BACE (M-83) (sc-10748;Santa Cruz Biotechnology).
Kim H.J., Chang K.A., Ha T.Y., Kim J., Ha S., Shin K.Y., Moon C., Nacken W., Kim H.S, & Suh Y.H. (2014). S100A9 Knockout Decreases the Memory Impairment and Neuropathology in Crossbreed Mice of Tg2576 and S100A9 Knockout Mice Model. PLoS ONE, 9(2), e88924.
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4

Protein Expression Analysis of Colonic Tissue

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Protein lysates were made from the colonic laminar propria scraping and were quantified using a Pierce BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA, USA). Proteins (100 μg) were loaded onto 12% Bis-Tris SDS NuPAGE gel (Invitrogen) followed by transfer onto PVDF transfer membrane (88518; Thermo Fisher Scientific). The membrane was then incubated with primary antibodies against F4/80 (ab6640; Abcam), S100A8 (AF3059; R&D Systems), occludin (ab31721; Abcam) or claudin 5 (ab15106; Abcam) overnight, followed by incubation with secondary antibodies conjugated to horseradish peroxidase, and developed with Pierce ECL substrate (32106; Thermo Fisher Scientific) using CL-XPosure film (34091; Thermo Fisher Scientific). Actin (69100; MP Biomedicals) was used as a loading control.
Kim Y.E., Lee M., Gu H., Kim J., Jeong S., Yeo S., Lee Y.J., Im S.H., Sung Y.C., Kim H.J., Weissman I.L, & Ahn G.O. (2018). HIF-1α activation in myeloid cells accelerates dextran sodium sulfate-induced colitis progression in mice. Disease Models & Mechanisms, 11(7), dmm033241.
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5

Oral Carcinogenesis Induction in Mice

Cited 2 times
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To induce oral carcinogenesis, 4-NQO (50 μg/ml ad libitum) was added to the water of C57BL/6j wild-type (N = 12) and S100A8/A9null mice (N = 12) for 16 weeks as described [37 (link), 38 (link)]. During weeks 17 to 22, animals received water without 4-NQO. Control mice (n=6 each genotype) received water without 4-NQO during the entire 22-week experiment. All animals were euthanized at 22 weeks. The tongue, maxillary gingiva, palate, and buccal mucosae were harvested for histopathologic examination using H/E staining. Intraoral epithelial dysplastic and/or carcinomatous lesions from 4-NQO-treated wild-type and S100A8/A9null mice (N = 7 in each group) were immunostained for 53BP1 (Abcam, ab36823, 1:200 dil.) and S100A8 (R&D, AF3059, 1:500 dil.) as described [35 (link)]. Staining for S100A8 was used as a surrogate for S100A8/A9. At least 2 microscopic lesions per animal were selected and 20X-magnification photomicrographs per lesion were captured using the Leica DM6 B (Leica Microsystems CMS GmbH) microscope. Quantification of 53BP1 positive nuclei and statistical analysis were performed as above.
Argyris P.P., Saavedra F., Malz C., Stone I.A., Wei Y., Boyle W.S., Johnstone K.F., Khammanivong A, & Herzberg M.C. (2023). Intracellular Calprotectin (S100A8/A9) Facilitates DNA Damage Responses and Promotes Apoptosis in Head and Neck Squamous Cell Carcinoma. Oral oncology, 137, 106304.
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