polyclonal antibody against s100a8  (Proteintech)

Journal: Bone Research

Article Title: Specific inflammatory osteoclast precursors induced during chronic inflammation give rise to highly active osteoclasts associated with inflammatory bone loss

doi: 10.1038/s41413-022-00206-z

Figure Lengend Snippet: S100A8/A9 proteins augment differentiation and osteoclastic activity derived from iOCPs but not from hOCPs. a Expression of S100A8, S100A9 and RAGE in sorted control and inflamed BM iOCPs and hOCPs. The protein phosphatase 2A catalytic subunit (PP2Ac) is shown as a loading control. b Densitometry measurements of three biological repeats (no statistical analysis presented). c Sorted control and inflamed BM iOCPs and hOCPs (5 × 10 4 ) were cultured on the Osteo assay surface with or without anti-RAGE blocking antibodies (bar: 50 µm). d Pit area quantitation. e Sorted iOCPs and hOCPs (5 × 10 4 ) from the BM of control mice were cultured on the Osteo assay surface in combination with a recombinant S100A8/A9 heterodimer and anti-RAGE blocking antibodies as indicated (bar: 50 µm). f Pit area quantitation. c – f Depict representative results for two independent experiments, n = 5 for each group. Line: median, box: 25th-75th percentile, whiskers: range. * P < 0.05 (Mann–Whitney test and Holm multiplicity correction)

Article Snippet: For the detection of S100A8/A9, membranes were blocked in 10% skim milk in PBST and probed with goat polyclonal antibodies against S100A8 (R&D, Cat#: AF3059) and S100A9 (R&D, Cat#: AF2065), followed by incubation with peroxidase-conjugated rabbit anti-goat IgG (Jackson ImmunoResearch Labs, Cat#: 305–035–045).

Techniques: Activity Assay, Derivative Assay, Expressing, Control, Cell Culture, Blocking Assay, Quantitation Assay, Recombinant, MANN-WHITNEY

Journal: Bone Research

Article Title: Specific inflammatory osteoclast precursors induced during chronic inflammation give rise to highly active osteoclasts associated with inflammatory bone loss

doi: 10.1038/s41413-022-00206-z

Figure Lengend Snippet: TNF-α ablation abrogates IBL by targeting iOCPs but not the hOCP response during chronic inflammation. a Expression of S100a8 and S100a9 transcripts in the right tibia of control and inflamed WT and tnf - α −/− mice. b iOCP and hOCP frequencies and absolute numbers in the BM of control and inflamed WT and tnf - α −/− mice (left panel) and the fractions in the peripheral blood (right panel). c . Sorted iOCPs and hOCPs (5 × 10 4 )- from the BM of inflamed WT and tnf - α −/− mice cultured on the Osteo assay surface (bar: 50 µm). d Quantitation of total pit area/well. e Representative 3D modeling from femoral microCT scans of control and inflamed WT and tnf - -α −/− mice. f Cortical cross-sectional area fraction (Ct.Ar/Tt.Ar) and trabecular BV/TV. g Morphometric indices; Ct.Ar, Tb.N, Tb.Sp and Tb.Th. h Expression of Rankl , Opg and M-csf in right tibiae. a , b and e – h WT: control n = 7, inflamed n = 7; tnf - α −/− : control n = 8, inflamed n = 8; representative results for three independent experiments (Mann–Whitney test and Bonferroni multiplicity correction for a and f – h , Holm multiplicity correction for b . c , d: n = 8 for each group, representative results for three independent experiments (Mann–Whitney test and Holm multiplicity correction). Line: median, box: 25th–75th percentile, whiskers: range. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.000 1

Article Snippet: For the detection of S100A8/A9, membranes were blocked in 10% skim milk in PBST and probed with goat polyclonal antibodies against S100A8 (R&D, Cat#: AF3059) and S100A9 (R&D, Cat#: AF2065), followed by incubation with peroxidase-conjugated rabbit anti-goat IgG (Jackson ImmunoResearch Labs, Cat#: 305–035–045).

Techniques: Expressing, Control, Cell Culture, Quantitation Assay, MANN-WHITNEY

Journal: Oncogene

Article Title: Loss of Keratin 17 induces tissue-specific cytokine polarization and cellular differentiation in HPV16-driven cervical tumorigenesis in vivo

doi: 10.1038/onc.2016.102

Figure Lengend Snippet: Differentiation of cervical epithelia is induced in the absence of K17. (a) Normalized expression for S100a8 and S100a9 gene transcripts in cervical (left) and ear (right) tissue of HPV16tg/+ and HPV16tg/+;Krt17−/− mice at 3 months post implantation. (b) Immunoblots for S100a8 in cervical (top) and ear (bottom) tissues of WT, HPV16tg/+ and HPV16tg/+;Krt17−/− mice at 3 months post implantation. Each lane represents a distinct biological replicate. Actin serves as loading control. Densitometric quantification below. (c) Normalized expression for differentiation markers in cervical tissue of HPV16tg/+ and HPV16tg/+;Krt17−/− mice at 1 month (left) and 3 months (right) post implantation. (d) Immunoblots for K1 and filaggrin in cervical tissue of HPV16tg/+ and HPV16tg/+;Krt17−/− mice at 1 month post implantation. Each lane represents a distinct biological replicate. Actin serves as loading control. Densitometric quantification below. (e, f) Immunostaining for K1 in (e) transformation zone epithelium and (f) ear skin epidermis of HPV16tg/+ and HPV16tg/+;Krt17−/− mice at 1 month post implantation. Nuclei are stained with DAPI (blue). Dotted line separates epithelium (above line) from underlying stroma (below line). Bar = 100 μm. All error bars are s.e.m. *P<0.05. NS; no significance.

Article Snippet: Antibodies Primary antibodies used included rabbit polyclonal antisera directed against K17, 41 K14 (Covance #AF64, Princeton, NJ, USA), K1 (Biolegend #AF109, San Diego, CA, USA), phospho-histone H3 (Cell Signaling #9701, Danvers, MA, USA), CDKN2A (Thermo Scientific #MA1-16664, Waltham, MA, USA) and filaggrin (Biolegend #PRB-417P); goat polyclonal antisera against S100A8 (Santa Cruz #SC-8113, Dallas, TX, USA); rat monoclonal antibodies against CD11b (eBiosciences #11-0112-81, San Diego, CA, USA), F4/80 (AbD Serotec #MCA497, Raleigh, NC, USA) and CD4 (BD Biosciences #550280, San Jose, CA, USA); and mouse monoclonal antisera against HPV E7 (Invitrogen #28-0006), PECAM-1 (Chemicon #CBL1337, Billerica, MA, USA), p63α (Genetex #GTX23239, Irvine, CA, USA) and β-actin (Sigma #A5441, St Louis, MO, USA).

Techniques: Expressing, Western Blot, Control, Immunostaining, Transformation Assay, Staining