Structured Review

Santa Cruz Biotechnology goat
Goat, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/goat/product/Santa Cruz Biotechnology
Average 93 stars, based on 18 article reviews
Price from $9.99 to $1999.99
goat - by Bioz Stars, 2020-09
93/100 stars

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Produced:

Article Title: SULF2 overexpression positively regulates tumorigenicity of human prostate cancer cells
Article Snippet: .. After 3 days, cells were fixed in methanol:acetone (1:1) for 2 min and incubated with primary antibody anti-SULF2 (H-80, Santa Cruz Biotechnology, CA, USA), polyclonal anti-human vimentin produced in goat (Santa Cruz Biotechnology, CA, USA), monoclonal anti-human-β-catenin produced in mouse (MAB13291-100, R & D Systems, MA, USA); Alexa 594 conjugated phalloidin (Invitrogen, Life Technologies Corporation, CA, USA) in PBS containing 5% FBS for 1 h. Subsequently, cells were incubated with secondary antibody conjugated with a fluorescent marker diluted 1:200 in PBS for 40 min in the dark. .. Cell nuclei were stained with DAPI 1:1000 in PBS with 0.01% saponin for 30 min.

Concentration Assay:

Article Title: Orexin Neurons Express a Functional Pancreatic Polypeptide Y4 Receptor
Article Snippet: .. Sections were then incubated in a mixture of primary antibodies, including the c-Fos antibody made in the rabbit (SC-52; Santa Cruz Biotechnology) (used at concentration of 1:25,000), the orexin antibody made in the goat (Santa Cruz Biotechnology) (used at a concentration of 1:10,000), and the MCH antibody made in the chicken (Bachem, Torrance, CA) (used at a concentration of 1:3000) for 48 hr at 4°C. .. After washes in KPBS, tissue was incubated for 1 hr in biotinylated donkey anti-rabbit antibody (1:600; Jackson ImmunoResearch) and then washed and incubated in avidin–biotin solution (Vectastain) for 1 hr. c-Fos-IR was visualized with the chromagen 3,3′-diaminobenzidine enhanced with nickel chloride.

Article Title: Disruption of sphingolipid metabolism augments ceramide-induced autophagy in preeclampsia
Article Snippet: .. For negative controls, the primary antibody was replaced by a corresponding concentration of nonimmune mouse (Santa Cruz Biotechnology, sc-2025), goat (Santa Cruz Biotechnology, sc-2028) or rabbit IgG. ..

Incubation:

Article Title: Orexin Neurons Express a Functional Pancreatic Polypeptide Y4 Receptor
Article Snippet: .. Sections were then incubated in a mixture of primary antibodies, including the c-Fos antibody made in the rabbit (SC-52; Santa Cruz Biotechnology) (used at concentration of 1:25,000), the orexin antibody made in the goat (Santa Cruz Biotechnology) (used at a concentration of 1:10,000), and the MCH antibody made in the chicken (Bachem, Torrance, CA) (used at a concentration of 1:3000) for 48 hr at 4°C. .. After washes in KPBS, tissue was incubated for 1 hr in biotinylated donkey anti-rabbit antibody (1:600; Jackson ImmunoResearch) and then washed and incubated in avidin–biotin solution (Vectastain) for 1 hr. c-Fos-IR was visualized with the chromagen 3,3′-diaminobenzidine enhanced with nickel chloride.

Article Title: SULF2 overexpression positively regulates tumorigenicity of human prostate cancer cells
Article Snippet: .. After 3 days, cells were fixed in methanol:acetone (1:1) for 2 min and incubated with primary antibody anti-SULF2 (H-80, Santa Cruz Biotechnology, CA, USA), polyclonal anti-human vimentin produced in goat (Santa Cruz Biotechnology, CA, USA), monoclonal anti-human-β-catenin produced in mouse (MAB13291-100, R & D Systems, MA, USA); Alexa 594 conjugated phalloidin (Invitrogen, Life Technologies Corporation, CA, USA) in PBS containing 5% FBS for 1 h. Subsequently, cells were incubated with secondary antibody conjugated with a fluorescent marker diluted 1:200 in PBS for 40 min in the dark. .. Cell nuclei were stained with DAPI 1:1000 in PBS with 0.01% saponin for 30 min.

Article Title: A Circadian Clock in the Olfactory Bulb Anticipates Feeding during Food Anticipatory Activity
Article Snippet: .. Free floating sections were incubated in PER1 antibody raised in goat (sc-7724, Santa Cruz Biotechnology, Santa Cruz, CA, USA) or in c-Fos antibody raised in goat (sc-52G, Santa Cruz Biotechnology, Santa Cruz, CA, USA) both diluted at 1∶2000, in 3% normal horse serum and 0.3% Triton X-100 (Sigma, St. Louis MO, USA), for 48 h at 4°C. .. It was followed by incubation in secondary antibody, biotinylated horse anti-goat diluted at 1∶200 (Vector Laboratories, Burlingame, CA, USA) in PB and 0.3% Triton X-100 for 1 h at room temperature.

Marker:

Article Title: SULF2 overexpression positively regulates tumorigenicity of human prostate cancer cells
Article Snippet: .. After 3 days, cells were fixed in methanol:acetone (1:1) for 2 min and incubated with primary antibody anti-SULF2 (H-80, Santa Cruz Biotechnology, CA, USA), polyclonal anti-human vimentin produced in goat (Santa Cruz Biotechnology, CA, USA), monoclonal anti-human-β-catenin produced in mouse (MAB13291-100, R & D Systems, MA, USA); Alexa 594 conjugated phalloidin (Invitrogen, Life Technologies Corporation, CA, USA) in PBS containing 5% FBS for 1 h. Subsequently, cells were incubated with secondary antibody conjugated with a fluorescent marker diluted 1:200 in PBS for 40 min in the dark. .. Cell nuclei were stained with DAPI 1:1000 in PBS with 0.01% saponin for 30 min.

other:

Article Title: The Soluble Recombinant Neisseria meningitidis Adhesin NadAΔ351–405 Stimulates Human Monocytes by Binding to Extracellular Hsp90
Article Snippet: Antibodies Rabbit anti-hsp90(H114), against peptide 610–723 of C-terminal human hsp90, goat anti-hsp90(N17), against N-terminal region of human hsp90, and goat anti-hsp70(K20), against a C-terminal peptide of human hsp70, antibodies (Abs) were obtained from Santa Cruz Biotechnology.

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  • 92
    Santa Cruz Biotechnology goat anti s100a8
    <t>S100A8/A9</t> expression in PASMCs of patients with PAH. A. Immunohistochemical staining of S100A8 (green) (left figures), α-SMA (red) (center figures) and merge images (right figures) in distal pulmonary arteries of patients with IPAH (upper figures), patients with HPAH (middle figures) and without PAH (non-PAH) (lower figures). B. Immunohistochemical staining of S100A9 (green) (left figures), α-SMA (red) (center figures) and merge images (right figures) in distal pulmonary arteries of patients with IPAH (upper figures), patients with HPAH (middle figures) and patients without PAH (non-PAH) (lower figures). C. Immunocytochemical analysis of S100A8 (green) (left figures), S100A9 (green) (center figures) and negative control staining without 1 st antibody (left figures) in cultured PASMCs of patients with IPAH (upper figures), patients with HPAH (middle figures) and patients without PAH (non-PAH) (lower figures). Nuclear staining was performed by use of DAPI (blue). Bar = 100 μm.
    Goat Anti S100a8, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti s100a8/product/Santa Cruz Biotechnology
    Average 92 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    goat anti s100a8 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    85
    Santa Cruz Biotechnology goat anti irf8
    Cross-regulation of <t>IRF8</t> and PU.1 in microglial activation . (A) PU.1 directly targets IRF8 expression in microglial activation. IRF8 and PU.1 target sites at the gene locus of Irf8 are shown. (B and C) PU.1 regulates IRF8 expression. Cx3cr1 +/+ ; Pu.1 fl / fl vs. Cx3cr1 CreERT2 /+ ; Pu.1 fl / fl mice were treated with 4-hydroxytamoxifen to induce the Cre-recombinase activity and then subjected to optic nerve injury. The control and injured nerves were processed for the whole-tissue immunolabeling of IRF8 or PU.1 and imaged on the lightsheet microscope. (B) Representative orthogonal 3D-projection images of the optic nerves. (C) Density of IRF8 + or PU.1 + nuclei. n = 3, mean ± SEM, * P
    Goat Anti Irf8, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti irf8/product/Santa Cruz Biotechnology
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti irf8 - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    93
    Santa Cruz Biotechnology goat anti rsp5
    Yca1 maintains proteostasis through direct participation in the ubiquitin cycle. Phosphorylation of the S346 residue (P) on Yca1 by candidate kinases such as casein kinase 1 (CK1), is recognized by <t>Rsp5,</t> which leads to ubiquitination (U) at K355. In turn, these modifications influence Yca1’s ability to limit protein aggregation and autophagy as well as target and liberate free ubiquitin from precursor proteins (Rps31). Once liberated, the free ubiquitin may cycle back to maintain Yca1 activity or be used for modifying other proteins to maintain proteostasis within the cell
    Goat Anti Rsp5, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti rsp5/product/Santa Cruz Biotechnology
    Average 93 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    goat anti rsp5 - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    92
    Santa Cruz Biotechnology goat anti mouse fractalkine antibody
    IP 3 -mediated calcium signaling participates in <t>fractalkine-induced</t> [Ca 2+ ] i release in BV-2 cells. ( A ) [Ca 2+ ] i was increased by fractalkine in the media with calcium. ( B ) [Ca 2+ ] i was increased by fractalkine in the media with calcium-free. ( C ) The elevation of [Ca 2+ ] i by fractalkine was inhibited by 2-APB. * P
    Goat Anti Mouse Fractalkine Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti mouse fractalkine antibody/product/Santa Cruz Biotechnology
    Average 92 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse fractalkine antibody - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    Image Search Results


    S100A8/A9 expression in PASMCs of patients with PAH. A. Immunohistochemical staining of S100A8 (green) (left figures), α-SMA (red) (center figures) and merge images (right figures) in distal pulmonary arteries of patients with IPAH (upper figures), patients with HPAH (middle figures) and without PAH (non-PAH) (lower figures). B. Immunohistochemical staining of S100A9 (green) (left figures), α-SMA (red) (center figures) and merge images (right figures) in distal pulmonary arteries of patients with IPAH (upper figures), patients with HPAH (middle figures) and patients without PAH (non-PAH) (lower figures). C. Immunocytochemical analysis of S100A8 (green) (left figures), S100A9 (green) (center figures) and negative control staining without 1 st antibody (left figures) in cultured PASMCs of patients with IPAH (upper figures), patients with HPAH (middle figures) and patients without PAH (non-PAH) (lower figures). Nuclear staining was performed by use of DAPI (blue). Bar = 100 μm.

    Journal: PLoS ONE

    Article Title: Crucial role of RAGE in inappropriate increase of smooth muscle cells from patients with pulmonary arterial hypertension

    doi: 10.1371/journal.pone.0203046

    Figure Lengend Snippet: S100A8/A9 expression in PASMCs of patients with PAH. A. Immunohistochemical staining of S100A8 (green) (left figures), α-SMA (red) (center figures) and merge images (right figures) in distal pulmonary arteries of patients with IPAH (upper figures), patients with HPAH (middle figures) and without PAH (non-PAH) (lower figures). B. Immunohistochemical staining of S100A9 (green) (left figures), α-SMA (red) (center figures) and merge images (right figures) in distal pulmonary arteries of patients with IPAH (upper figures), patients with HPAH (middle figures) and patients without PAH (non-PAH) (lower figures). C. Immunocytochemical analysis of S100A8 (green) (left figures), S100A9 (green) (center figures) and negative control staining without 1 st antibody (left figures) in cultured PASMCs of patients with IPAH (upper figures), patients with HPAH (middle figures) and patients without PAH (non-PAH) (lower figures). Nuclear staining was performed by use of DAPI (blue). Bar = 100 μm.

    Article Snippet: Rabbit anti-human RAGE antibody (Santa Cruz Biotechnology), mouse anti-αSMA antibody (Sigma-Aldrich), goat anti-S100A8 (calgranulin A) antibody (Santa Cruz Biotechnology) and goat anti-S100A9 (calgranulin B) antibody (Santa Cruz Biotechnology) were used.

    Techniques: Expressing, Immunohistochemistry, Staining, Negative Control, Cell Culture

    Cross-regulation of IRF8 and PU.1 in microglial activation . (A) PU.1 directly targets IRF8 expression in microglial activation. IRF8 and PU.1 target sites at the gene locus of Irf8 are shown. (B and C) PU.1 regulates IRF8 expression. Cx3cr1 +/+ ; Pu.1 fl / fl vs. Cx3cr1 CreERT2 /+ ; Pu.1 fl / fl mice were treated with 4-hydroxytamoxifen to induce the Cre-recombinase activity and then subjected to optic nerve injury. The control and injured nerves were processed for the whole-tissue immunolabeling of IRF8 or PU.1 and imaged on the lightsheet microscope. (B) Representative orthogonal 3D-projection images of the optic nerves. (C) Density of IRF8 + or PU.1 + nuclei. n = 3, mean ± SEM, * P

    Journal: Protein & Cell

    Article Title: Transcriptional mechanism of IRF8 and PU.1 governs microglial activation in neurodegenerative condition

    doi: 10.1007/s13238-018-0599-3

    Figure Lengend Snippet: Cross-regulation of IRF8 and PU.1 in microglial activation . (A) PU.1 directly targets IRF8 expression in microglial activation. IRF8 and PU.1 target sites at the gene locus of Irf8 are shown. (B and C) PU.1 regulates IRF8 expression. Cx3cr1 +/+ ; Pu.1 fl / fl vs. Cx3cr1 CreERT2 /+ ; Pu.1 fl / fl mice were treated with 4-hydroxytamoxifen to induce the Cre-recombinase activity and then subjected to optic nerve injury. The control and injured nerves were processed for the whole-tissue immunolabeling of IRF8 or PU.1 and imaged on the lightsheet microscope. (B) Representative orthogonal 3D-projection images of the optic nerves. (C) Density of IRF8 + or PU.1 + nuclei. n = 3, mean ± SEM, * P

    Article Snippet: Antibodies Primary antibodies used in the experiments were rat anti-PECAM1 (BD Biosciences #553370, RRID:AB_394816), rat anti-LYVE1 (Thermo Fisher Scientific #14-0443-82, RRID:AB_1633414), chicken anti-GFP (Aves Labs #GFP-1010, RRID:AB_2307313), rat anti-CD11b (Biolegend #101202, RRID:AB_312785), rabbit anti-RFP (Rockland #600-401-379, RRID:AB_2209751), mouse anti-Iba1 (Millipore #MABN92, RRID:AB_10917271), rabbit anti-Olig2 (Millipore #AB9610, RRID:AB_10141047), goat anti-IRF8 (Santa Cruz #sc-6058, RRID:AB_649510), rabbit anti-PU.1 (Cell Signaling #2258, RRID:AB_10693421), rabbit anti-Ki67 (Millipore #AB9260, RRID:AB_2142366) and mouse anti-p-IκBα (Cell Signaling #9246, RRID:AB_2267145).

    Techniques: Activation Assay, Expressing, Mouse Assay, Activity Assay, Immunolabeling, Microscopy

    Transcription factor IRF8 acts in microglial activation . (A and B) Microglial activation depends on Sarm1-mediated neurodegeneration. Sarm1 +/+ vs. Sarm1 −/− mice were subjected to optic nerve injury. The control and injured nerves were processed for the whole-tissue immunolabeling of CD11b and imaged on the lightsheet microscope. (A) Representative orthogonal 3D-projection images of the injured nerves. (B) Density of CD11b + microglia. n = 4, mean ± SEM, * P

    Journal: Protein & Cell

    Article Title: Transcriptional mechanism of IRF8 and PU.1 governs microglial activation in neurodegenerative condition

    doi: 10.1007/s13238-018-0599-3

    Figure Lengend Snippet: Transcription factor IRF8 acts in microglial activation . (A and B) Microglial activation depends on Sarm1-mediated neurodegeneration. Sarm1 +/+ vs. Sarm1 −/− mice were subjected to optic nerve injury. The control and injured nerves were processed for the whole-tissue immunolabeling of CD11b and imaged on the lightsheet microscope. (A) Representative orthogonal 3D-projection images of the injured nerves. (B) Density of CD11b + microglia. n = 4, mean ± SEM, * P

    Article Snippet: Antibodies Primary antibodies used in the experiments were rat anti-PECAM1 (BD Biosciences #553370, RRID:AB_394816), rat anti-LYVE1 (Thermo Fisher Scientific #14-0443-82, RRID:AB_1633414), chicken anti-GFP (Aves Labs #GFP-1010, RRID:AB_2307313), rat anti-CD11b (Biolegend #101202, RRID:AB_312785), rabbit anti-RFP (Rockland #600-401-379, RRID:AB_2209751), mouse anti-Iba1 (Millipore #MABN92, RRID:AB_10917271), rabbit anti-Olig2 (Millipore #AB9610, RRID:AB_10141047), goat anti-IRF8 (Santa Cruz #sc-6058, RRID:AB_649510), rabbit anti-PU.1 (Cell Signaling #2258, RRID:AB_10693421), rabbit anti-Ki67 (Millipore #AB9260, RRID:AB_2142366) and mouse anti-p-IκBα (Cell Signaling #9246, RRID:AB_2267145).

    Techniques: Activation Assay, Mouse Assay, Immunolabeling, Microscopy

    IRF8 and PU.1 cooperatively target microglial activation-related genes . (A) IRF8 and PU.1 cooperatively regulate microglial activation-related genes. The genes that contained the IRF8-PU.1 co-target sites were subjected to GO enrichment analysis. Enriched biological processes and signaling pathways are shown. (B) Activation of the NF-κB pathway in microglial activation. Wild-type mice were subjected to optic nerve injury. The control and injured nerves were examined by immunohistochemistry for p-IκBα and CD11b. (C) IRF8 and PU.1 directly target the key signaling components of the NF-κb pathway. IRF8 and PU.1 target sites at the indicated gene loci are shown. The sequence of the DNA probe derived from the IRF8-PU.1 co-target site at each locus is included, with the IRF-ETS composite motif highlighted. (D and E) Biochemical assembly of the ternary complex of the composite-motif DNA with IRF8 and PU.1 proteins. (D) Synergetic binding of IRF8 and PU.1 to the composite-motif DNA, derived from the gene loci of Ripk2 , Tak1 , Ikbkb or Nfkb1 , was examined by EMSA. (E) Supershift of the composite-motif DNA/IRF8/PU.1 ternary complex with anti-IRF8 or anti-PU.1 antibody

    Journal: Protein & Cell

    Article Title: Transcriptional mechanism of IRF8 and PU.1 governs microglial activation in neurodegenerative condition

    doi: 10.1007/s13238-018-0599-3

    Figure Lengend Snippet: IRF8 and PU.1 cooperatively target microglial activation-related genes . (A) IRF8 and PU.1 cooperatively regulate microglial activation-related genes. The genes that contained the IRF8-PU.1 co-target sites were subjected to GO enrichment analysis. Enriched biological processes and signaling pathways are shown. (B) Activation of the NF-κB pathway in microglial activation. Wild-type mice were subjected to optic nerve injury. The control and injured nerves were examined by immunohistochemistry for p-IκBα and CD11b. (C) IRF8 and PU.1 directly target the key signaling components of the NF-κb pathway. IRF8 and PU.1 target sites at the indicated gene loci are shown. The sequence of the DNA probe derived from the IRF8-PU.1 co-target site at each locus is included, with the IRF-ETS composite motif highlighted. (D and E) Biochemical assembly of the ternary complex of the composite-motif DNA with IRF8 and PU.1 proteins. (D) Synergetic binding of IRF8 and PU.1 to the composite-motif DNA, derived from the gene loci of Ripk2 , Tak1 , Ikbkb or Nfkb1 , was examined by EMSA. (E) Supershift of the composite-motif DNA/IRF8/PU.1 ternary complex with anti-IRF8 or anti-PU.1 antibody

    Article Snippet: Antibodies Primary antibodies used in the experiments were rat anti-PECAM1 (BD Biosciences #553370, RRID:AB_394816), rat anti-LYVE1 (Thermo Fisher Scientific #14-0443-82, RRID:AB_1633414), chicken anti-GFP (Aves Labs #GFP-1010, RRID:AB_2307313), rat anti-CD11b (Biolegend #101202, RRID:AB_312785), rabbit anti-RFP (Rockland #600-401-379, RRID:AB_2209751), mouse anti-Iba1 (Millipore #MABN92, RRID:AB_10917271), rabbit anti-Olig2 (Millipore #AB9610, RRID:AB_10141047), goat anti-IRF8 (Santa Cruz #sc-6058, RRID:AB_649510), rabbit anti-PU.1 (Cell Signaling #2258, RRID:AB_10693421), rabbit anti-Ki67 (Millipore #AB9260, RRID:AB_2142366) and mouse anti-p-IκBα (Cell Signaling #9246, RRID:AB_2267145).

    Techniques: Activation Assay, Mouse Assay, Immunohistochemistry, Sequencing, Derivative Assay, Binding Assay

    Post-developmental IRF8 is required for microglial activation . (A) IRF8 and PU.1 cooperatively target microglial activation-related genes. IRF8 and PU.1 target sites at the indicated gene loci are shown. (B–G) Post-developmental IRF8 is required for microglial activation. Cx3cr1 +/+ ; Irf8 fl / fl vs. Cx3cr1 CreERT2 /+ ; Irf8 fl / fl mice were treated with 4-hydroxytamoxifen to induce the Cre-recombinase activity and then subjected to optic nerve injury. (B) Expression levels of the indicated genes in the optic nerves were examined by qPCR analysis. n = 4, mean ± SEM, * P

    Journal: Protein & Cell

    Article Title: Transcriptional mechanism of IRF8 and PU.1 governs microglial activation in neurodegenerative condition

    doi: 10.1007/s13238-018-0599-3

    Figure Lengend Snippet: Post-developmental IRF8 is required for microglial activation . (A) IRF8 and PU.1 cooperatively target microglial activation-related genes. IRF8 and PU.1 target sites at the indicated gene loci are shown. (B–G) Post-developmental IRF8 is required for microglial activation. Cx3cr1 +/+ ; Irf8 fl / fl vs. Cx3cr1 CreERT2 /+ ; Irf8 fl / fl mice were treated with 4-hydroxytamoxifen to induce the Cre-recombinase activity and then subjected to optic nerve injury. (B) Expression levels of the indicated genes in the optic nerves were examined by qPCR analysis. n = 4, mean ± SEM, * P

    Article Snippet: Antibodies Primary antibodies used in the experiments were rat anti-PECAM1 (BD Biosciences #553370, RRID:AB_394816), rat anti-LYVE1 (Thermo Fisher Scientific #14-0443-82, RRID:AB_1633414), chicken anti-GFP (Aves Labs #GFP-1010, RRID:AB_2307313), rat anti-CD11b (Biolegend #101202, RRID:AB_312785), rabbit anti-RFP (Rockland #600-401-379, RRID:AB_2209751), mouse anti-Iba1 (Millipore #MABN92, RRID:AB_10917271), rabbit anti-Olig2 (Millipore #AB9610, RRID:AB_10141047), goat anti-IRF8 (Santa Cruz #sc-6058, RRID:AB_649510), rabbit anti-PU.1 (Cell Signaling #2258, RRID:AB_10693421), rabbit anti-Ki67 (Millipore #AB9260, RRID:AB_2142366) and mouse anti-p-IκBα (Cell Signaling #9246, RRID:AB_2267145).

    Techniques: Activation Assay, Mouse Assay, Activity Assay, Expressing, Real-time Polymerase Chain Reaction

    Yca1 maintains proteostasis through direct participation in the ubiquitin cycle. Phosphorylation of the S346 residue (P) on Yca1 by candidate kinases such as casein kinase 1 (CK1), is recognized by Rsp5, which leads to ubiquitination (U) at K355. In turn, these modifications influence Yca1’s ability to limit protein aggregation and autophagy as well as target and liberate free ubiquitin from precursor proteins (Rps31). Once liberated, the free ubiquitin may cycle back to maintain Yca1 activity or be used for modifying other proteins to maintain proteostasis within the cell

    Journal: Cell Discovery

    Article Title: The metacaspase Yca1 maintains proteostasis through multiple interactions with the ubiquitin system

    doi: 10.1038/s41421-018-0071-9

    Figure Lengend Snippet: Yca1 maintains proteostasis through direct participation in the ubiquitin cycle. Phosphorylation of the S346 residue (P) on Yca1 by candidate kinases such as casein kinase 1 (CK1), is recognized by Rsp5, which leads to ubiquitination (U) at K355. In turn, these modifications influence Yca1’s ability to limit protein aggregation and autophagy as well as target and liberate free ubiquitin from precursor proteins (Rps31). Once liberated, the free ubiquitin may cycle back to maintain Yca1 activity or be used for modifying other proteins to maintain proteostasis within the cell

    Article Snippet: The goat anti-Rsp5 (sc-26193; Santa Cruz Biotechnology, TX, USA) polyclonal antibody was used to detect presence of Rsp5.

    Techniques: Activity Assay

    Yca1 interacts with components of the ubiquitin pathway. a LC-MS/MS analysis of Yca1-RFP pulldown depicted ubiquitin (Ubi4) and the E3 ligase Rsp5 in high abundance (Total Spectra Counts). Data represented as mean ± SEM. n = 3. b Yca1-GFP cells were used to isolate ubiquitinated proteins which was further probed with anti-GFP antibody to detect the presence of Yca1-GFP. The arrows highlight the full length and processed forms of Yca1-GFP. Predicted size of full length Yca1-GFP is 76 kDa and processed form in 37 kDa. Protein G beads and anti-IgG antibody conjugated beads were used as controls. Lowercase letters denote independent replicates. n = 3. c Extracts in b were probed with anti-ubiquitin antibody to detect the presence of the bait. The asterisk (*) highlights the presence of IgG. d Rsp5-RFP was expressed from a plasmid in the Yca1-GFP strain and assessed for interaction between Yca1 and Rsp5 via anti-RFP immunoprecipitation followed by anti-GFP immunoblotting. Lowercase letters denote replicate samples. The arrows highlight the full length and processed forms of Yca1-GFP. The empty vector transformed Yca1-GFP cells (RFP) was used as an interaction control. Anti-IgG antibody conjugated beads were used to probe replicate ‘a’ as an additional control. n = 3. e Extracts in d were probed with anti-RFP antibody to detect the presence of the bait Rsp5-RFP (highlighted with an arrow). The estimated size of Rsp5-RFP is 119 kDa. The asterisk highlights the presence of IgG

    Journal: Cell Discovery

    Article Title: The metacaspase Yca1 maintains proteostasis through multiple interactions with the ubiquitin system

    doi: 10.1038/s41421-018-0071-9

    Figure Lengend Snippet: Yca1 interacts with components of the ubiquitin pathway. a LC-MS/MS analysis of Yca1-RFP pulldown depicted ubiquitin (Ubi4) and the E3 ligase Rsp5 in high abundance (Total Spectra Counts). Data represented as mean ± SEM. n = 3. b Yca1-GFP cells were used to isolate ubiquitinated proteins which was further probed with anti-GFP antibody to detect the presence of Yca1-GFP. The arrows highlight the full length and processed forms of Yca1-GFP. Predicted size of full length Yca1-GFP is 76 kDa and processed form in 37 kDa. Protein G beads and anti-IgG antibody conjugated beads were used as controls. Lowercase letters denote independent replicates. n = 3. c Extracts in b were probed with anti-ubiquitin antibody to detect the presence of the bait. The asterisk (*) highlights the presence of IgG. d Rsp5-RFP was expressed from a plasmid in the Yca1-GFP strain and assessed for interaction between Yca1 and Rsp5 via anti-RFP immunoprecipitation followed by anti-GFP immunoblotting. Lowercase letters denote replicate samples. The arrows highlight the full length and processed forms of Yca1-GFP. The empty vector transformed Yca1-GFP cells (RFP) was used as an interaction control. Anti-IgG antibody conjugated beads were used to probe replicate ‘a’ as an additional control. n = 3. e Extracts in d were probed with anti-RFP antibody to detect the presence of the bait Rsp5-RFP (highlighted with an arrow). The estimated size of Rsp5-RFP is 119 kDa. The asterisk highlights the presence of IgG

    Article Snippet: The goat anti-Rsp5 (sc-26193; Santa Cruz Biotechnology, TX, USA) polyclonal antibody was used to detect presence of Rsp5.

    Techniques: Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Plasmid Preparation, Immunoprecipitation, Transformation Assay

    Ubiquitination at K355 is mediated by S346 dependent on the interaction with Rsp5. a Proteins interacting with FL, K355A, S346A and RFP (vector control) were isolated and probed for presence of ubiquitin and ubiquitin conjugate material with anti-ubiquitin antibody. n = 3. Ub – highlights ubiquitin bands and IgG H/L highlights immunoglobin heavy and light chains. b Extracts from a were probed for the presence of Rsp5 using anti-Rsp5 antibody. The arrow highlights the position of Rsp5. The expected size of Rsp5 is 92 kDa. ‘ns’ identifies non-specific bands. n = 3. c Extracts from a were probed for the presence of the respective fusion proteins via anti-RFP immunoblotting. The arrows highlight the full length and processed from Yca1-RFP. The expected size of the full length fusion proteins is 76 kDa. The expected sizes of the processed forms of the fusion proteins are 63 kDa and 37 kDa. IgG H/L highlights immunoglobin heavy and light chains. n = 3. A shorter exposure for this blot is depicted in Supplementary Fig. S1c , which was used for densitometry and normalization. d Graph depicting the level of interaction between FL, K355A and S346A and ubiquitin and ubiquitinated material as assessed via densitometry, normalized to the amount of bait (individual RFP fusion) recovered in c (sum of bands indicated with arrows). n = 3. Data represented as mean ± SEM. * p value

    Journal: Cell Discovery

    Article Title: The metacaspase Yca1 maintains proteostasis through multiple interactions with the ubiquitin system

    doi: 10.1038/s41421-018-0071-9

    Figure Lengend Snippet: Ubiquitination at K355 is mediated by S346 dependent on the interaction with Rsp5. a Proteins interacting with FL, K355A, S346A and RFP (vector control) were isolated and probed for presence of ubiquitin and ubiquitin conjugate material with anti-ubiquitin antibody. n = 3. Ub – highlights ubiquitin bands and IgG H/L highlights immunoglobin heavy and light chains. b Extracts from a were probed for the presence of Rsp5 using anti-Rsp5 antibody. The arrow highlights the position of Rsp5. The expected size of Rsp5 is 92 kDa. ‘ns’ identifies non-specific bands. n = 3. c Extracts from a were probed for the presence of the respective fusion proteins via anti-RFP immunoblotting. The arrows highlight the full length and processed from Yca1-RFP. The expected size of the full length fusion proteins is 76 kDa. The expected sizes of the processed forms of the fusion proteins are 63 kDa and 37 kDa. IgG H/L highlights immunoglobin heavy and light chains. n = 3. A shorter exposure for this blot is depicted in Supplementary Fig. S1c , which was used for densitometry and normalization. d Graph depicting the level of interaction between FL, K355A and S346A and ubiquitin and ubiquitinated material as assessed via densitometry, normalized to the amount of bait (individual RFP fusion) recovered in c (sum of bands indicated with arrows). n = 3. Data represented as mean ± SEM. * p value

    Article Snippet: The goat anti-Rsp5 (sc-26193; Santa Cruz Biotechnology, TX, USA) polyclonal antibody was used to detect presence of Rsp5.

    Techniques: Plasmid Preparation, Isolation

    IP 3 -mediated calcium signaling participates in fractalkine-induced [Ca 2+ ] i release in BV-2 cells. ( A ) [Ca 2+ ] i was increased by fractalkine in the media with calcium. ( B ) [Ca 2+ ] i was increased by fractalkine in the media with calcium-free. ( C ) The elevation of [Ca 2+ ] i by fractalkine was inhibited by 2-APB. * P

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: IP3-Mediated Calcium Signaling Is Involved in the Mechanism of Fractalkine-Induced Hyperalgesia Response

    doi: 10.12659/MSM.913787

    Figure Lengend Snippet: IP 3 -mediated calcium signaling participates in fractalkine-induced [Ca 2+ ] i release in BV-2 cells. ( A ) [Ca 2+ ] i was increased by fractalkine in the media with calcium. ( B ) [Ca 2+ ] i was increased by fractalkine in the media with calcium-free. ( C ) The elevation of [Ca 2+ ] i by fractalkine was inhibited by 2-APB. * P

    Article Snippet: Sections were analyzed by immunofluorescence as previously described using goat anti-mouse fractalkine antibody (1: 50; SC-7227, Santa Cruz Biotechnology, Dallas, TX), rabbit anti-mouse Iba-1 (1: 500; Wako Chemicals, Richmond, VA), and rabbit anti-mouse GFAP (1: 50; ZSGB-BIO, Beijing, China) with secondary FITC-labeled anti-rabbit IgG Ab (1: 50; Proteintech, San Diego, CA) or TRITC-labeled anti-goat IgG Ab (1: 50; Proteintech).

    Techniques:

    Fractalkine injection lead to thermal hyperalgesia and activated microglia in vivo . ( A ) The thermal nociceptive threshold of mice after receiving fractalkine, 2-APB, antiCX3CR1, and SB203580. ( B ) The immunofluorescence of FKN (fractalkine), Iba-1, and GFAP in mice brain tissues without or with fractalkine. * P

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: IP3-Mediated Calcium Signaling Is Involved in the Mechanism of Fractalkine-Induced Hyperalgesia Response

    doi: 10.12659/MSM.913787

    Figure Lengend Snippet: Fractalkine injection lead to thermal hyperalgesia and activated microglia in vivo . ( A ) The thermal nociceptive threshold of mice after receiving fractalkine, 2-APB, antiCX3CR1, and SB203580. ( B ) The immunofluorescence of FKN (fractalkine), Iba-1, and GFAP in mice brain tissues without or with fractalkine. * P

    Article Snippet: Sections were analyzed by immunofluorescence as previously described using goat anti-mouse fractalkine antibody (1: 50; SC-7227, Santa Cruz Biotechnology, Dallas, TX), rabbit anti-mouse Iba-1 (1: 500; Wako Chemicals, Richmond, VA), and rabbit anti-mouse GFAP (1: 50; ZSGB-BIO, Beijing, China) with secondary FITC-labeled anti-rabbit IgG Ab (1: 50; Proteintech, San Diego, CA) or TRITC-labeled anti-goat IgG Ab (1: 50; Proteintech).

    Techniques: Injection, In Vivo, Mouse Assay, Immunofluorescence

    Fractalkine upregulated pro-inflammatory cytokines in vivo. ( A, B ) The increase of IL-1β and TNF-α by treatment with fractalkine in RT-PCR analysis. ( C, D ) The increase of IL-1β and TNF-α by treatment with fractalkine in ELISA analysis. ( E, F ) The decrease of IL-1β and TNF-α by treatment with anti-CX3CR1, 2-APB, and SB203580. * P

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: IP3-Mediated Calcium Signaling Is Involved in the Mechanism of Fractalkine-Induced Hyperalgesia Response

    doi: 10.12659/MSM.913787

    Figure Lengend Snippet: Fractalkine upregulated pro-inflammatory cytokines in vivo. ( A, B ) The increase of IL-1β and TNF-α by treatment with fractalkine in RT-PCR analysis. ( C, D ) The increase of IL-1β and TNF-α by treatment with fractalkine in ELISA analysis. ( E, F ) The decrease of IL-1β and TNF-α by treatment with anti-CX3CR1, 2-APB, and SB203580. * P

    Article Snippet: Sections were analyzed by immunofluorescence as previously described using goat anti-mouse fractalkine antibody (1: 50; SC-7227, Santa Cruz Biotechnology, Dallas, TX), rabbit anti-mouse Iba-1 (1: 500; Wako Chemicals, Richmond, VA), and rabbit anti-mouse GFAP (1: 50; ZSGB-BIO, Beijing, China) with secondary FITC-labeled anti-rabbit IgG Ab (1: 50; Proteintech, San Diego, CA) or TRITC-labeled anti-goat IgG Ab (1: 50; Proteintech).

    Techniques: In Vivo, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    Fractalkine upregulated p-p38MAPK protein in vivo. ( A ) The p-p38MAPK protein was increased after treatment with fractalkine. ( B ) The p-p38MAPK protein was attenuated by pretreatment with anti-CX3CR1, 2-APB, and SB203580. * P

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: IP3-Mediated Calcium Signaling Is Involved in the Mechanism of Fractalkine-Induced Hyperalgesia Response

    doi: 10.12659/MSM.913787

    Figure Lengend Snippet: Fractalkine upregulated p-p38MAPK protein in vivo. ( A ) The p-p38MAPK protein was increased after treatment with fractalkine. ( B ) The p-p38MAPK protein was attenuated by pretreatment with anti-CX3CR1, 2-APB, and SB203580. * P

    Article Snippet: Sections were analyzed by immunofluorescence as previously described using goat anti-mouse fractalkine antibody (1: 50; SC-7227, Santa Cruz Biotechnology, Dallas, TX), rabbit anti-mouse Iba-1 (1: 500; Wako Chemicals, Richmond, VA), and rabbit anti-mouse GFAP (1: 50; ZSGB-BIO, Beijing, China) with secondary FITC-labeled anti-rabbit IgG Ab (1: 50; Proteintech, San Diego, CA) or TRITC-labeled anti-goat IgG Ab (1: 50; Proteintech).

    Techniques: In Vivo

    IP 3 -mediated calcium signaling is involved in fractalkine-induced inflammatory responses in vitro. ( A, B ) The increased of IL-1β and TNF-α by exposed to fractalkine persistently. ( C, D ) The increased of IL-1β and TNF-α was decreased by 2-APB. ( E, F ) The mRNA of IL-1β and TNF-α were increased by exposed to fractalkine persistently. ( G, H ) The mRNA of IL-1β and TNF-α were decreased by 2-APB. * P

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: IP3-Mediated Calcium Signaling Is Involved in the Mechanism of Fractalkine-Induced Hyperalgesia Response

    doi: 10.12659/MSM.913787

    Figure Lengend Snippet: IP 3 -mediated calcium signaling is involved in fractalkine-induced inflammatory responses in vitro. ( A, B ) The increased of IL-1β and TNF-α by exposed to fractalkine persistently. ( C, D ) The increased of IL-1β and TNF-α was decreased by 2-APB. ( E, F ) The mRNA of IL-1β and TNF-α were increased by exposed to fractalkine persistently. ( G, H ) The mRNA of IL-1β and TNF-α were decreased by 2-APB. * P

    Article Snippet: Sections were analyzed by immunofluorescence as previously described using goat anti-mouse fractalkine antibody (1: 50; SC-7227, Santa Cruz Biotechnology, Dallas, TX), rabbit anti-mouse Iba-1 (1: 500; Wako Chemicals, Richmond, VA), and rabbit anti-mouse GFAP (1: 50; ZSGB-BIO, Beijing, China) with secondary FITC-labeled anti-rabbit IgG Ab (1: 50; Proteintech, San Diego, CA) or TRITC-labeled anti-goat IgG Ab (1: 50; Proteintech).

    Techniques: In Vitro