rabbit polyclonal anti-human full-length (fl)rankl antibody sc-9073  (Santa Cruz Biotechnology)

Journal: International journal of medical sciences

Article Title: Leptin regulates OPG and RANKL expression in Gingival Fibroblasts and Tissues of Chronic Periodontitis Patients.

doi: 10.7150/ijms.56151

Figure Lengend Snippet: Figure 1. Immunohistochemical staining of Leptin, OPG, and RANKL in control and chronic periodontitis groups. A-C. Immunohistochemical staining of Leptin in the control group(A), the moderate chronic periodontitis group (B), and the severe chronic periodontitis group (C). D-F. Immunohistochemical staining of OPG in the control group (D), the moderate chronic periodontitis group (E), and the severe chronic periodontitis group (F). G-I. Immunohistochemical staining of RANKL in the control group (G), the moderate chronic periodontitis group (H), and the severe chronic periodontitis group (I). Red arrows indicate the positive cells. EL: epithelial layer, LP: lamina propria.

Article Snippet: Leptin Rabbit Anti-Human Polyclonal Antibody (CSB-PA009805, CUSABIO, China), OPG Rabbit AntiHuman Polyclonal Antibody (CSB-PA003597, CUSABIO, China), RANKL Rabbit Anti-Human Polyclonal Antibody (CSB-PA023986LA01HU, CUSABIO, China), DAB (Servicebio, China), Dulbecco’s Modified Eagle Medium (DMEM), Fetal Bovine Serum (Hyclone, America), Trypsin (Gibco, America), TRizol, PrimeScript RT reagent kit, SYBR Green mix (CWBIO, China) were used.

Techniques: Immunohistochemical staining, Staining, Control

Journal: International journal of medical sciences

Article Title: Leptin regulates OPG and RANKL expression in Gingival Fibroblasts and Tissues of Chronic Periodontitis Patients.

doi: 10.7150/ijms.56151

Figure Lengend Snippet: Figure 2. RANKL and OPG expression levels in leptin-treated HGF. A. Immunohistochemical staining of vimentin in HGF. B. Immunohistochemical staining of keratin in HGF. C. Expression of OPG, RANKL and OPG/RANKL at mRNA level in different leptin concentration groups (ug/ml).

Article Snippet: Leptin Rabbit Anti-Human Polyclonal Antibody (CSB-PA009805, CUSABIO, China), OPG Rabbit AntiHuman Polyclonal Antibody (CSB-PA003597, CUSABIO, China), RANKL Rabbit Anti-Human Polyclonal Antibody (CSB-PA023986LA01HU, CUSABIO, China), DAB (Servicebio, China), Dulbecco’s Modified Eagle Medium (DMEM), Fetal Bovine Serum (Hyclone, America), Trypsin (Gibco, America), TRizol, PrimeScript RT reagent kit, SYBR Green mix (CWBIO, China) were used.

Techniques: Expressing, Immunohistochemical staining, Staining, Concentration Assay

Journal: Genes and immunity

Article Title: Activated Human T Cells Express Alternative mRNA Transcripts Encoding a Secreted Form of RANKL

doi: 10.1038/gene.2013.29

Figure Lengend Snippet: A. Color coded schematics showing the exon structure for the human TNFSF11 mRNA transcripts aligned with the known RANKL protein domains. Key: E, Exon; EST, expressed sequence tag; IC, intracellular; TM, transmembrane; EC, extracellular; A +1 TG, translation start site. B. Schematic showing the genomic structure of the human TNFSF11/RANKL locus. Numbering corresponds to the human genomic contig Accession: AL139382. C. Vista genome alignment of the human TNFSF11 genomic locus (identified by exon-intron schematic at top) to rhesus monkey, dog and mouse genomic loci. Shaded areas identify regions where sequence identity across a 50 bp window is 70% or greater. Key: Light blue shading : conservation of untranslated sequences; dark blue shading : conserved coding sequence; pink shading : conserved intron sequence. Location of repeat elements and single nucleotide polymorphisms (SNPs) are marked as indicated in the figure.

Article Snippet: The eluted protein was subjected to SDS-PAGE followed by Western blot analysis with a rabbit anti-human RANKL polyclonal antibody (PeproTech Inc, Rocky Hill NJ, USA).

Techniques: Sequencing

Journal: Genes and immunity

Article Title: Activated Human T Cells Express Alternative mRNA Transcripts Encoding a Secreted Form of RANKL

doi: 10.1038/gene.2013.29

Figure Lengend Snippet: A. Semi-quantitative PCR followed by southern hybridization using primers to E1B and E5, shows expression of TNFSF11 Variant 2 in Saos-2 and Jurkat T cells. A second PCR product containing E1B and E2-5 was also amplified. Expression of HPRT is shown as a loading control. B. Western blots showing that the longest open reading frame of TNFSF11 Variant 2 is translated and secreted when expressed as a myc-fusion protein in 293FT cells. Identity of the protein in cell lysates and culture media was confirmed using both an anti-RANKL antibody and anti-myc antibody. C. and D. Conditioned media from 293FT cells transfected with secRANKL vector supports osteoclast differentiation (identified by counting TRAP-positive cells (red staining) with > 3 nuclei which is blocked by inclusion of 200ng/mL OPG. C. Representative results are shown C. Osteoclast quantitation/well (average of 3 wells +/−SD). D. Representative images of each culture condition (the equivalent of 10ng/mL RANKL is shown for secRANKL vector).

Article Snippet: The eluted protein was subjected to SDS-PAGE followed by Western blot analysis with a rabbit anti-human RANKL polyclonal antibody (PeproTech Inc, Rocky Hill NJ, USA).

Techniques: Real-time Polymerase Chain Reaction, Hybridization, Expressing, Variant Assay, Amplification, Control, Western Blot, Transfection, Plasmid Preparation, Staining, Quantitation Assay

Journal: Genes and immunity

Article Title: Activated Human T Cells Express Alternative mRNA Transcripts Encoding a Secreted Form of RANKL

doi: 10.1038/gene.2013.29

Figure Lengend Snippet: A. TNFSF11 transcript expression in Jurkat T cells stimulated for 24 hrs with PMA/ionomycin (P/I) with and without cyclosporine A (CsA) pre-treatment. Left panel: representative semi-quantitative RT-PCR results using primers specific for TNFSF11 Variant 2 (E1B-E5) (minimum of 3 biologic repeats). GAPDH is shown as loading control. Right panel: quantitative RT-PCR showing relative expression (GAPDH house keeping gene) of total TNFSF11 expression (n = 3). * p ≤ 0.05. B. TNFSF11 mRNA expression in primary human T cells stimulated for 6 hrs with either anti-CD2/anti-CD3/anti-CD28 conjugated to MACS Bead particles (MACS-ABMB) or P/I with and without CsA pre-treatment. Left panel: representative semi-quantitative RT-PCR using primers specific for TNFSF11 Variant 1 (amplified from E1D-E4) and Transcript Variant 2 (amplified from E1B-E5) (minimum of 3 biologic repeats). GAPDH was used as the housekeeping gene. Right panel: quantitative RT-PCR showing relative expression (GAPDH housekeeping gene) of total TNFSF11 and TNFSF11 Variant 1 expression (n = 3). * and # indicate statistically significant differences (p ≤ 0.05) between indicated conditions for TNFSF11 Variant 1 and total TNFSF11 expression respectively.

Article Snippet: The eluted protein was subjected to SDS-PAGE followed by Western blot analysis with a rabbit anti-human RANKL polyclonal antibody (PeproTech Inc, Rocky Hill NJ, USA).

Techniques: Expressing, Quantitative RT-PCR, Variant Assay, Control, Amplification

Journal: Genes and immunity

Article Title: Activated Human T Cells Express Alternative mRNA Transcripts Encoding a Secreted Form of RANKL

doi: 10.1038/gene.2013.29

Figure Lengend Snippet: A. Western Bot showing expression of secreted RANKL protein isoform in Jurkat T cell lysates and culture media with or without PMA/ionomycin (P/I) stimulation. RANKL-GST (a recombinant full-length transmembrane RANKL protein) was run as a control. Cont – control. B. Flow cytometry analysis of surface (membrane-bound) RANKL protein expression (i) Unstimulated MG-63 and (ii) Jurkat T cells, vehicle control or P/I stimulated. C. Flow cytometry analysis of intracellular RANKL protein expression in permeabilized Jurkat T cells treated with or without brefeldin before harvest. (i) Control unstimulated or (ii) PMA/ionomycin stimulated Jurkat T cells without brefeldin treatment (iii) Control unstimulated or (iv) P/I stimulated Jurkat T cells treated with brefeldin.

Article Snippet: The eluted protein was subjected to SDS-PAGE followed by Western blot analysis with a rabbit anti-human RANKL polyclonal antibody (PeproTech Inc, Rocky Hill NJ, USA).

Techniques: Western Blot, Expressing, Recombinant, Control, Flow Cytometry, Membrane

Journal: Genes and immunity

Article Title: Activated Human T Cells Express Alternative mRNA Transcripts Encoding a Secreted Form of RANKL

doi: 10.1038/gene.2013.29

Figure Lengend Snippet: Primers

Article Snippet: The eluted protein was subjected to SDS-PAGE followed by Western blot analysis with a rabbit anti-human RANKL polyclonal antibody (PeproTech Inc, Rocky Hill NJ, USA).

Techniques: Sequencing, Variant Assay