elisa kit (Thermo Fisher)

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Ethylene Glycol

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Thermo Scientific Pierce Ethylene Glycol Solution provides exceptional antifreeze protection and storage stability for antibody enzyme conjugates because it is purified to remove impurities commonly found traditional glycerol stocks Features of Ethylene Glycol • Specially purified to remove impurities such as aldehydes peroxides iron and UV absorbing hydrocarbons • Suitable for enzyme storage without the worry of losing enzymatic activity • Stable for months This product is a 50 w v aqueous solution of highly purified ethylene glycol When mixed in equal volume with purified protein samples such as primary antibodies the solution stabilizes and maintains the mixture as a liquid during freezer storage 20° Ethylene glycol is a suitable alternative to glycerol for most protein storage applications In fact this preparation of ethylene glycol is devoid of various common glycerol impurities and potiential degradation products that are damaging to certain enzymes

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29810

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Structured Review

Thermo Fisher elisa kit

TSG-6 secreted by canine adipose tissue-derived mesenchymal stem cells (cAT-MSCs) inhibits inflammatory response and apoptosis in the colon. a Levels of tumor necrosis factor <t>(TNF)-α,</t> interleukin (IL)-6, and IL-10 in colons were assessed by <t>ELISA.</t> b Representative immunofluorescence staining of colon tissue sections using annexin-V antibody or propidium iodide (PI), and the percentage of the annexin-V-positive cells are shown. Four to six mice per group were used. Results are shown as the mean ± standard deviation. * P

elisa kit - by Bioz Stars, 2021-03

86/100 stars

Images

1) Product Images from "TSG-6 released from intraperitoneally injected canine adipose tissue-derived mesenchymal stem cells ameliorate inflammatory bowel disease by inducing M2 macrophage switch in mice"

Article Title: TSG-6 released from intraperitoneally injected canine adipose tissue-derived mesenchymal stem cells ameliorate inflammatory bowel disease by inducing M2 macrophage switch in mice

Journal: Stem Cell Research & Therapy

doi: 10.1186/s13287-018-0841-1

Figure Legend Snippet: TSG-6 secreted by canine adipose tissue-derived mesenchymal stem cells (cAT-MSCs) inhibits inflammatory response and apoptosis in the colon. a Levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-10 in colons were assessed by ELISA. b Representative immunofluorescence staining of colon tissue sections using annexin-V antibody or propidium iodide (PI), and the percentage of the annexin-V-positive cells are shown. Four to six mice per group were used. Results are shown as the mean ± standard deviation. * P

Techniques Used: Derivative Assay, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Mouse Assay, Standard Deviation

2) Product Images from "Mefunidone Attenuates Tubulointerstitial Fibrosis in a Rat Model of Unilateral Ureteral Obstruction"

Article Title: Mefunidone Attenuates Tubulointerstitial Fibrosis in a Rat Model of Unilateral Ureteral Obstruction

Journal: PLoS ONE

doi: 10.1371/journal.pone.0129283

Mefunidone inhibited cytokine release and affected ERK1/2, NF-κB and STAT3 signaling pathways in HK-2 cells and macrophages. A . Mefunidone inhibited cytokine release in HK-2 cells culture supernatants using enzyme-linked immunosorbent assay (ELISA). B . Mefunidone inhibited cytokine release in macrophages culture supernatants using ELISA. C and E . Mefunidone affected the phosphorylation of ERK1/2, NF-κB and STAT3 stimulated with TNF-α in HK-2 cells. The inhibitor was PD98059 (10 −6 μM), BAY (10μM) or AG490 (150μM) respectively. The columnar section was the optical density of the gels tested by Western blot. D and G . Mefunidone affected the phosphorylation of ERK1/2, NF-κB and STAT3 stimulated with LPS in macrophages. The inhibitor was PD98059 (10 −6 μM), BAY (10μM) or AG490 (150μM) respectively. The columnar section was the optical density of the gels tested by Western blot. F . Assays of luciferase activity were performed and pNF-κB-luciferase activity was normalized to pRL-SV40 luciferase activity in HK-2 cells. The data were presented as means±SD, n = 3. (*p

Figure Legend Snippet: Mefunidone inhibited cytokine release and affected ERK1/2, NF-κB and STAT3 signaling pathways in HK-2 cells and macrophages. A . Mefunidone inhibited cytokine release in HK-2 cells culture supernatants using enzyme-linked immunosorbent assay (ELISA). B . Mefunidone inhibited cytokine release in macrophages culture supernatants using ELISA. C and E . Mefunidone affected the phosphorylation of ERK1/2, NF-κB and STAT3 stimulated with TNF-α in HK-2 cells. The inhibitor was PD98059 (10 −6 μM), BAY (10μM) or AG490 (150μM) respectively. The columnar section was the optical density of the gels tested by Western blot. D and G . Mefunidone affected the phosphorylation of ERK1/2, NF-κB and STAT3 stimulated with LPS in macrophages. The inhibitor was PD98059 (10 −6 μM), BAY (10μM) or AG490 (150μM) respectively. The columnar section was the optical density of the gels tested by Western blot. F . Assays of luciferase activity were performed and pNF-κB-luciferase activity was normalized to pRL-SV40 luciferase activity in HK-2 cells. The data were presented as means±SD, n = 3. (*p

Techniques Used: Enzyme-linked Immunosorbent Assay, Western Blot, Luciferase, Activity Assay

3) Product Images from "Effective Pro-Inflammatory Induced Activity of GALT, a Conserved Antigen in A. Pleuropneumoniae, Improves the Cytokines Secretion of Macrophage via p38, ERK1/2 and JNK MAPKs Signal Pathway"

Article Title: Effective Pro-Inflammatory Induced Activity of GALT, a Conserved Antigen in A. Pleuropneumoniae, Improves the Cytokines Secretion of Macrophage via p38, ERK1/2 and JNK MAPKs Signal Pathway

Journal: Frontiers in Cellular and Infection Microbiology

doi: 10.3389/fcimb.2018.00337

Pro-inflammatory cytokines induced by GALT. Induction of cytokines in Raw 264.7 macrophages by recombinant GALT stimulation. Raw 264.7 macrophages were incubated with 10 μg/mL GALT and 200 ng/mL LPS (positive control) for 12 h, as well as single culture media (negative control). Protein levels of TNF-α, IL-1β, IL-6 in the culture supernatants were determined by ELISA.

Figure Legend Snippet: Pro-inflammatory cytokines induced by GALT. Induction of cytokines in Raw 264.7 macrophages by recombinant GALT stimulation. Raw 264.7 macrophages were incubated with 10 μg/mL GALT and 200 ng/mL LPS (positive control) for 12 h, as well as single culture media (negative control). Protein levels of TNF-α, IL-1β, IL-6 in the culture supernatants were determined by ELISA.

Techniques Used: Recombinant, Incubation, Positive Control, Negative Control, Enzyme-linked Immunosorbent Assay

4) Product Images from "(+)-JQ1 attenuated LPS-induced microglial inflammation via MAPK/NFκB signaling"

Article Title: (+)-JQ1 attenuated LPS-induced microglial inflammation via MAPK/NFκB signaling

Journal: Cell & Bioscience

doi: 10.1186/s13578-018-0258-7

(+)-JQ1 reduces expressions of LPS/Aβ-induced proinflammatory cytokines in microglia. a BV2 cells were pretreated with (+)-JQ1 or the vehicle (DMSO) at the indicated dose, followed by LPS (100 ng/ml) stimulation for 4 h. b Alternatively, BV2 cells were pretreated with either (+)-JQ1 (400 nM) or DMSO, followed by LPS stimulation for the indicated period. After incubation, the cells were collected and lysed for quantitative analysis of transcription levels of IL-1β, IL-6 and TNFα by qRT-PCR. c , d BV2 cells were pretreated with (+)-JQ1, DMSO, or minomycine (mino) at the indicated dose, followed by LPS (100 ng/ml) or Aβ (10 μM) stimulation for 24 h. IL-1β, IL-6 and TNFα levels in the culture medium were determined via ELISA. The data are presented as the mean ± SEM (n = 3, * p ≤ 0.05)

Figure Legend Snippet: (+)-JQ1 reduces expressions of LPS/Aβ-induced proinflammatory cytokines in microglia. a BV2 cells were pretreated with (+)-JQ1 or the vehicle (DMSO) at the indicated dose, followed by LPS (100 ng/ml) stimulation for 4 h. b Alternatively, BV2 cells were pretreated with either (+)-JQ1 (400 nM) or DMSO, followed by LPS stimulation for the indicated period. After incubation, the cells were collected and lysed for quantitative analysis of transcription levels of IL-1β, IL-6 and TNFα by qRT-PCR. c , d BV2 cells were pretreated with (+)-JQ1, DMSO, or minomycine (mino) at the indicated dose, followed by LPS (100 ng/ml) or Aβ (10 μM) stimulation for 24 h. IL-1β, IL-6 and TNFα levels in the culture medium were determined via ELISA. The data are presented as the mean ± SEM (n = 3, * p ≤ 0.05)

Techniques Used: Incubation, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

MAPK but not PI3 K signaling is targeted by (+)-JQ1. BV2 cells were pretreated with either (+)-JQ1 (400 nM) or DMSO for 30 min and subjected to LPS stimulation. The phosphorylation of ERK, p38, JNK ( a ) and Akt ( b ) as well as the total protein content, was analyzed by immunoblot. The relative intensity of a given band was normalized. Representative results from three independent experiments are depicted here. The relative intensity of a given band was normalized (n = 6). c BV2 cells were treated with LPS (100 ng/ml) or JQ1 (400 nM), together with or without U0126 (ERK inhibitor) or SB203580 (p38 inhibitor). 24 h later, cell culture medium were collected and IL-1β, IL-6 and TNFα were tested by ELISA (n = 3). The data are presented as the mean ± SEM (* p ≤ 0.05, ** p ≤ 0.01)

Figure Legend Snippet: MAPK but not PI3 K signaling is targeted by (+)-JQ1. BV2 cells were pretreated with either (+)-JQ1 (400 nM) or DMSO for 30 min and subjected to LPS stimulation. The phosphorylation of ERK, p38, JNK ( a ) and Akt ( b ) as well as the total protein content, was analyzed by immunoblot. The relative intensity of a given band was normalized. Representative results from three independent experiments are depicted here. The relative intensity of a given band was normalized (n = 6). c BV2 cells were treated with LPS (100 ng/ml) or JQ1 (400 nM), together with or without U0126 (ERK inhibitor) or SB203580 (p38 inhibitor). 24 h later, cell culture medium were collected and IL-1β, IL-6 and TNFα were tested by ELISA (n = 3). The data are presented as the mean ± SEM (* p ≤ 0.05, ** p ≤ 0.01)

Techniques Used: Cell Culture, Enzyme-linked Immunosorbent Assay

(+)-JQ1 attenuates NFκB phosphorylation, translocation and transcription induced by LPS. a BV2 cells were transfected with either pGL3-NFκB or pGL3 along with pRL-TK. After 24 h, the cells were treated with either DMSO or (+)-JQ1 (400 nM) followed by LPS (100 ng/ml) stimulation. NFκB-mediated promoter activity was assessed by a luciferase assay (n = 3). b p65 nuclear translocation upon LPS (100 ng/ml) stimulation in either the presence or absence of (+)-JQ1 (400 nM) was detected by immunofluorescence staining and confocal microscopy. p65 immunoreactivity is shown in red, and nuclei were stained with DAPI (blue). (Scale bar = 20 μm). c BV2 cells preincubated with either (+)-JQ1 (400 nM) or DMSO were stimulated with LPS (100 ng/ml) for the indicated periods. Phosphorylation of IKKα/β (p-IKKα/β), IκBα (p-IκBα) and p65 (p-p65), as well as the total protein content, were analyzed by immnoblot. Representative results from three independent experiments are depicted here. The relative intensity of a given band was normalized (n = 6). d BV2 cells were treated with LPS (100 ng/ml) or JQ1 (400 nM), together with or without PDTC (p65 inhibitor). 24 h later, cell culture medium were collected and IL-1β, IL-6 and TNFα were tested by ELISA (n = 3). The data are presented as the mean ± SEM (* p ≤ 0.05, ** p ≤ 0.01)

Figure Legend Snippet: (+)-JQ1 attenuates NFκB phosphorylation, translocation and transcription induced by LPS. a BV2 cells were transfected with either pGL3-NFκB or pGL3 along with pRL-TK. After 24 h, the cells were treated with either DMSO or (+)-JQ1 (400 nM) followed by LPS (100 ng/ml) stimulation. NFκB-mediated promoter activity was assessed by a luciferase assay (n = 3). b p65 nuclear translocation upon LPS (100 ng/ml) stimulation in either the presence or absence of (+)-JQ1 (400 nM) was detected by immunofluorescence staining and confocal microscopy. p65 immunoreactivity is shown in red, and nuclei were stained with DAPI (blue). (Scale bar = 20 μm). c BV2 cells preincubated with either (+)-JQ1 (400 nM) or DMSO were stimulated with LPS (100 ng/ml) for the indicated periods. Phosphorylation of IKKα/β (p-IKKα/β), IκBα (p-IκBα) and p65 (p-p65), as well as the total protein content, were analyzed by immnoblot. Representative results from three independent experiments are depicted here. The relative intensity of a given band was normalized (n = 6). d BV2 cells were treated with LPS (100 ng/ml) or JQ1 (400 nM), together with or without PDTC (p65 inhibitor). 24 h later, cell culture medium were collected and IL-1β, IL-6 and TNFα were tested by ELISA (n = 3). The data are presented as the mean ± SEM (* p ≤ 0.05, ** p ≤ 0.01)

Techniques Used: Translocation Assay, Transfection, Activity Assay, Luciferase, Immunofluorescence, Staining, Confocal Microscopy, Cell Culture, Enzyme-linked Immunosorbent Assay

5) Product Images from "Synthesis and biological evaluation of a novel class of curcumin analogs as anti-inflammatory agents for prevention and treatment of sepsis in mouse model"

Article Title: Synthesis and biological evaluation of a novel class of curcumin analogs as anti-inflammatory agents for prevention and treatment of sepsis in mouse model

Journal: Drug Design, Development and Therapy

doi: 10.2147/DDDT.S75862

AMAC compounds 3f , 3m , 4b , and 4d inhibited LPS-induced TNF-α and IL-6 release in MPMs in a dose-dependent manner. Notes: MPMs were plated at a density of 4×10 5 /plate for overnight in 37°C and 5% CO 2 . Cells were pretreated with specific compound at indicated concentrations for 2 hours, followed by LPS (0.5 μg/mL) treatment for 22 hours. The levels of TNF-α ( A ) or IL-6 ( B ) in the culture medium were measured by ELISA and were normalized to the total amount of protein. The bars represent percent TNF-α or IL-6 level as compared to the LPS control. Each bar represents mean ± SD of three independent experiments. Statistical significance relative to the LPS group was indicated, * P

Figure Legend Snippet: AMAC compounds 3f , 3m , 4b , and 4d inhibited LPS-induced TNF-α and IL-6 release in MPMs in a dose-dependent manner. Notes: MPMs were plated at a density of 4×10 5 /plate for overnight in 37°C and 5% CO 2 . Cells were pretreated with specific compound at indicated concentrations for 2 hours, followed by LPS (0.5 μg/mL) treatment for 22 hours. The levels of TNF-α ( A ) or IL-6 ( B ) in the culture medium were measured by ELISA and were normalized to the total amount of protein. The bars represent percent TNF-α or IL-6 level as compared to the LPS control. Each bar represents mean ± SD of three independent experiments. Statistical significance relative to the LPS group was indicated, * P

Techniques Used: Enzyme-linked Immunosorbent Assay

Synthesis scheme, chemical structures, and anti-inflammatory activities of AMACs ( 3a – f , 3i – j , 3m , and 4a – m ). Notes: MPM cells were pretreated with AMACs (10 μM) for 2 hours, and then treated with LPS (0.5 μg/mL) for 22 hours. TNF-α and IL-6 levels in the culture medium were measured by ELISA and were normalized to the total amount of protein. The percent inhibition of TNF-α and IL-6 is represented. * P

Figure Legend Snippet: Synthesis scheme, chemical structures, and anti-inflammatory activities of AMACs ( 3a – f , 3i – j , 3m , and 4a – m ). Notes: MPM cells were pretreated with AMACs (10 μM) for 2 hours, and then treated with LPS (0.5 μg/mL) for 22 hours. TNF-α and IL-6 levels in the culture medium were measured by ELISA and were normalized to the total amount of protein. The percent inhibition of TNF-α and IL-6 is represented. * P

Techniques Used: Enzyme-linked Immunosorbent Assay, Inhibition

6) Product Images from "ARHGAP24 ameliorates inflammatory response through inactivating Rac1/Akt/NF-κB pathway in acute pneumonia model of rat"

Article Title: ARHGAP24 ameliorates inflammatory response through inactivating Rac1/Akt/NF-κB pathway in acute pneumonia model of rat

Journal: Annals of Translational Medicine

doi: 10.21037/atm-20-5000

The effects of ARHGAP24 on the levels of inflammatory cytokines in the BALF of acute pneumonia rats. The pro-inflammatory level, including TNF-α, IL-1β, IL-6 of each group detected by ELISA assay. *, P

Figure Legend Snippet: The effects of ARHGAP24 on the levels of inflammatory cytokines in the BALF of acute pneumonia rats. The pro-inflammatory level, including TNF-α, IL-1β, IL-6 of each group detected by ELISA assay. *, P

Techniques Used: Enzyme-linked Immunosorbent Assay

7) Product Images from "A Toxoplasma dense granule protein, GRA24, modulates the early immune response to infection by promoting a direct and sustained host p38 MAPK activation"

Article Title: A Toxoplasma dense granule protein, GRA24, modulates the early immune response to infection by promoting a direct and sustained host p38 MAPK activation

Journal: The Journal of Experimental Medicine

doi: 10.1084/jem.20130103

GRA24 promotes chemokine secretion and contributes to the control of early parasite replication in vivo at the site of infection. (A) IL-12p40 cytokine production by uninfected (u.i.) or 24-h-infected (Pru ku80 Δgra24 or parental strains) BMDM measured using ELISA. Means of three independent experiments ± SD are shown (*, P

Figure Legend Snippet: GRA24 promotes chemokine secretion and contributes to the control of early parasite replication in vivo at the site of infection. (A) IL-12p40 cytokine production by uninfected (u.i.) or 24-h-infected (Pru ku80 Δgra24 or parental strains) BMDM measured using ELISA. Means of three independent experiments ± SD are shown (*, P

Techniques Used: In Vivo, Infection, Enzyme-linked Immunosorbent Assay

8) Product Images from "Inhibitory effect of Korean Red Ginseng on melanocyte proliferation and its possible implication in GM-CSF mediated signaling"

Article Title: Inhibitory effect of Korean Red Ginseng on melanocyte proliferation and its possible implication in GM-CSF mediated signaling

Journal: Journal of Ginseng Research

doi: 10.5142/jgr.2013.37.389

Effect of Korean Red Ginseng (KRG) on the expression of cytokines in UV-irradiated SP-1 keratinocytes. Before UV irradiation, SP-1 keratinocyte were treated with serum free Eagle’s Minimum Essential Medium (EMEM) containing total extract or saponin of KRG (SKRG) (10, 20, and 50 ppm). After 24 h, SP-1 keratinocytes were irradiated with UVB at a dose of 30 mJ/cm 2 . Immediately, the cells were treated with 2% newborn calf serum EMEM containing total extract or SKRG (10, 20, and 50 ppm). After 24 h, the conditioned media was measured by enzyme-linked immunosorbent assay as described in the Materials and Methods section. (A) Tumor necrosis factor (TNF)-α, (B) granulocyte macrophage colony-stimulating factor (GM-CSF), (C) interleukin (IL)-4, and (D) interferon (IFN)-γ. Data are shown as average±SEM, DMSO, dimethyl sulfoxide. * p

Figure Legend Snippet: Effect of Korean Red Ginseng (KRG) on the expression of cytokines in UV-irradiated SP-1 keratinocytes. Before UV irradiation, SP-1 keratinocyte were treated with serum free Eagle’s Minimum Essential Medium (EMEM) containing total extract or saponin of KRG (SKRG) (10, 20, and 50 ppm). After 24 h, SP-1 keratinocytes were irradiated with UVB at a dose of 30 mJ/cm 2 . Immediately, the cells were treated with 2% newborn calf serum EMEM containing total extract or SKRG (10, 20, and 50 ppm). After 24 h, the conditioned media was measured by enzyme-linked immunosorbent assay as described in the Materials and Methods section. (A) Tumor necrosis factor (TNF)-α, (B) granulocyte macrophage colony-stimulating factor (GM-CSF), (C) interleukin (IL)-4, and (D) interferon (IFN)-γ. Data are shown as average±SEM, DMSO, dimethyl sulfoxide. * p

Techniques Used: Expressing, Irradiation, Enzyme-linked Immunosorbent Assay

9) Product Images from "Anti-inflammatory effects of Zea mays L. husk extracts"

Article Title: Anti-inflammatory effects of Zea mays L. husk extracts

Journal: BMC Complementary and Alternative Medicine

doi: 10.1186/s12906-016-1284-9

Effects of ZMHE on eotaxin-1 gene expression. a The eotaxin-1 luciferase reporter vector was transfected into NIH/3 T3 cells and cultured for 24 h. The cells were pretreated with ZMHE for 1 h and then stimulated with IL-4. Luciferase activity was calculated against IL-4-unstimulated control. b Cells were pretreated with ZMHE for 1 h and then further incubated with IL-4 (50 ng/ml) for 24 h. Eotaxin-1 release was then determined using an ELISA. The results are mean ± standard deviation (SD) ( n = 3). P

Figure Legend Snippet: Effects of ZMHE on eotaxin-1 gene expression. a The eotaxin-1 luciferase reporter vector was transfected into NIH/3 T3 cells and cultured for 24 h. The cells were pretreated with ZMHE for 1 h and then stimulated with IL-4. Luciferase activity was calculated against IL-4-unstimulated control. b Cells were pretreated with ZMHE for 1 h and then further incubated with IL-4 (50 ng/ml) for 24 h. Eotaxin-1 release was then determined using an ELISA. The results are mean ± standard deviation (SD) ( n = 3). P

Techniques Used: Expressing, Luciferase, Plasmid Preparation, Transfection, Cell Culture, Activity Assay, Incubation, Enzyme-linked Immunosorbent Assay, Standard Deviation

Effects of ZMHE on the expression ICAM-1. RAW264.7 cells were pretreated with ZMHE for 1 h before stimulation with LPS (200 ng/ml). After 24 h of incubation, the concentrations of sICAM-1 in the culture medium were measured by ELISA. The results are mean ± standard deviation (SD) ( n = 3). P

Figure Legend Snippet: Effects of ZMHE on the expression ICAM-1. RAW264.7 cells were pretreated with ZMHE for 1 h before stimulation with LPS (200 ng/ml). After 24 h of incubation, the concentrations of sICAM-1 in the culture medium were measured by ELISA. The results are mean ± standard deviation (SD) ( n = 3). P

Techniques Used: Expressing, Incubation, Enzyme-linked Immunosorbent Assay, Standard Deviation

10) Product Images from "Hyperoxia-induced hypertrophy and ion channel remodeling in left ventricle"

Article Title: Hyperoxia-induced hypertrophy and ion channel remodeling in left ventricle

Journal: American Journal of Physiology - Heart and Circulatory Physiology

doi: 10.1152/ajpheart.00474.2012

Intracellular TNF-α expression in hyperoxic mice. The intracellular TNF-α expression was measured using ELISA, and values were expressed in picograms of TNF-α per microgram of total protein. The values on y -axis are means ± SE ( n = 3). The numbers above the bar represent the fold values compared with normoxia group. * P ≤ 0.05.

Figure Legend Snippet: Intracellular TNF-α expression in hyperoxic mice. The intracellular TNF-α expression was measured using ELISA, and values were expressed in picograms of TNF-α per microgram of total protein. The values on y -axis are means ± SE ( n = 3). The numbers above the bar represent the fold values compared with normoxia group. * P ≤ 0.05.

Techniques Used: Expressing, Mouse Assay, Enzyme-linked Immunosorbent Assay

11) Product Images from "GSAP Regulates Amyloid Beta Production through Modulation of Amyloid Precursor Protein Trafficking"

Article Title: GSAP Regulates Amyloid Beta Production through Modulation of Amyloid Precursor Protein Trafficking

Journal: bioRxiv

doi: 10.1101/2020.11.12.379313

Effect of GSAP knockdown on Aβ secretation and APP distribution. (a) Downregulation of Aβ40 and Aβ42 secretation by GSAP knockdown using control (CTRL) and GSAP siRNA in N2a-695 cells. Secreted Aβ40 and Aβ42 levels were measured by ELISA (mean±SEM; n=3). (b) Immunoblotting analysis of the total APP protein level following transfection with CTRL and GSAP siRNA (Left) and quantification (Right). (mean±SEM; n=5). (c) Representative images of subcellular localization of APP and Golgi in living N2a cells infected with a baculovirus expressing an RFP-Golgi marker, and then transfected with APP-GFP plasmids. (d) Colocalization coefficients m1 quantification for APP-GFP and RFP-Golgi (n = 12; Scale bar: 10 μm). (e) Cell surface APP was biotinylated and precipitated with streptavidin-coupled beads following transfection with CTRL and GSAP siRNA. Total APP in the lysates and the precipitated surface APP were detected by immunoblotting (Left) and quantification (Right) (means±SEM; n=3). *P

Figure Legend Snippet: Effect of GSAP knockdown on Aβ secretation and APP distribution. (a) Downregulation of Aβ40 and Aβ42 secretation by GSAP knockdown using control (CTRL) and GSAP siRNA in N2a-695 cells. Secreted Aβ40 and Aβ42 levels were measured by ELISA (mean±SEM; n=3). (b) Immunoblotting analysis of the total APP protein level following transfection with CTRL and GSAP siRNA (Left) and quantification (Right). (mean±SEM; n=5). (c) Representative images of subcellular localization of APP and Golgi in living N2a cells infected with a baculovirus expressing an RFP-Golgi marker, and then transfected with APP-GFP plasmids. (d) Colocalization coefficients m1 quantification for APP-GFP and RFP-Golgi (n = 12; Scale bar: 10 μm). (e) Cell surface APP was biotinylated and precipitated with streptavidin-coupled beads following transfection with CTRL and GSAP siRNA. Total APP in the lysates and the precipitated surface APP were detected by immunoblotting (Left) and quantification (Right) (means±SEM; n=3). *P

Techniques Used: Enzyme-linked Immunosorbent Assay, Transfection, Infection, Expressing, Marker

Construct:

Article Title: Poleward transport of Eg5 by dynein-dynactin in Xenopus laevis egg extract spindles
Article Snippet: A sequence corresponding to amino acids 174–505 that are predicted to form the first coiled coil (cc1; ) of X. laevis p150Glued was amplified from a p150 X. laevis cDNA plasmid (RZPD) and inserted (EcoRI–XhoI) into pGEX-6P-1 (GE Healthcare). .. Protein purifications Full-length Eg5 without GFP and fluorescent Eg5 constructs were expressed in Sf9 insect cells using the Bac-to-Bac baculovirus system (Invitrogen). .. Cells from a 350-ml culture were harvested 72 h after infection, suspended in 3.5 ml of lysis buffer (50 mM KPi , pH 8.0, 250 mM KCl, 10 mM imidazole, 0.5 mM MgATP, 0.1% Triton X100, 5 mM mercaptoethanol [ME], and complete EDTA-free protease inhibitors [PI]; Roche) and frozen in liquid nitrogen.

BAC Assay:

Expressing:

Article Title: Regulation of inflammation and fibrosis by macrophages in lymphedema
Article Snippet: Scar index was calculated by comparing the ratio of orange/red to yellow/green staining and is expressed as an arbitrary number with higher numbers representing more fibrosis. .. VEGF-C protein expression was quantified in total protein isolated from the tail tissues using ELISA (eBiosciences) as previously described ( ). .. Briefly, skin and subcutaneous tissues were harvested and lysed using the Qiagen DNA/RNA/Protein mini kit using the manufacturer's methods (Qiagen, Valencia, CA).

Article Title: The grapevine kinome: annotation, classification and expression patterns in developmental processes and stress responses
Article Snippet: The expression data of 1,109 available grapevine kinase genes were collected and normalized. .. The gene expression data of 231 available kinase genes in response to salt, polyethylene glycol (PEG), cold, drought, heat and abscisic acid (ABA) treatments were downloaded from four selected Affymetrix datasets, GSE31594, GSE31677, GSE31675, and GSE31662. .. These datasets were used to analyze the expression of grapevine PKs in response to different stress treatments.

Article Title: Allele-specific siRNA silencing for the common Keratin 12 founder mutation in Meesmann epithelial corneal dystrophy
Article Snippet: All PCR reactions were carried out using GoTaq Flexi DNA polymerase system (Promega, Southampton, UK) according to manufacturer’s instructions using the following cycling conditions: 94°C for 4 min, then 32 cycles of 95°C for 30s, 62°C for 30s and 72°C for 15s, followed by 5 min at 72°C and a 4°C hold. .. 3.5 × 105 AD293 cells were seeded in 2ml DMEM (10% FBS) in a 6 well plate, incubated for 24h and transfected with 1μg of plasmid DNA expressing K12-Arg135Thr-eGFP fusion protein using 3.3μl Lipofectamine 2000 (Invitrogen, Paisley, UK) in Optimem serum-free medium following manufacturer’s instructions. .. After 24h the medium was replaced and the cells transfected with siRNA K12-Arg135Thr-5 to a final concentration of 3nM using 6μl of Lipofectamine RNAiMAX (Invitrogen, Paisley, UK) according to the manufacturer’s instructions for 6h.

Isolation:

Enzyme-linked Immunosorbent Assay:

Article Title: Effects of continuous positive airway pressure on exhaled transforming growth factor-β and vascular endothelial growth factor in patients with obstructive sleep apnea
Article Snippet: The liquid component (serum) was immediately transferred into a clean polypropylene tube using a Pasteur pipette. .. The TGF-β and VEGF protein levels in the serum and EBC were determined by enzyme-linked immunosorbent assay (ELISA) with Human TGF-β and VEGF Instant ELISA kits (eBioscience, San Diego, CA, USA). ..

other:

Article Title: Crowder-Induced Conformational Ensemble Shift in Escherichia coli Prolyl-tRNA Synthetase
Article Snippet: Materials and Methods All crowding agents were purchased from Sigma-Aldrich (St. Louis, MO), except for polyethylene glycol (PEG) 8000 (Thermo Fisher Scientific, Waltham, MA).

Article Title: Cross-linked Polystyrene Sulfonic Acid and Polyethylene Glycol as a Low-fouling Material
Article Snippet: 75,000 g/mol Polystyrene sulfonic acid (PSS) was purchased from Sigma; 20,000 g/mol Polyethylene glycol (PEG) was purchased from Alfa Aesar.