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    Structured Review

    ALPCO elisa kit
    EREG stimulates leptin secretion and lipolysis in mice with diet-induced obesity (DIO). (A) DIO WT male mice (n=7/ group) were injected with 100μL PBS (Veh) with and without EREG (1.5ng/g body weight or 20ng/per iAb depot) into both epididymal iAb fat pads every other day for 2 weeks. Total mRNA was isolated from one whole iAb fad pad. Expression of <t>Lep</t> was measured using NanoString assay. Normalized data represent mean±SD, n=5; Mann-Whitney U test. (B) Expression of LEP protein levels were measured in plasma in the same mice group by <t>ELISA.</t> Data (mean±SD, n=5/group). Independent Student’s t -test. (C) Non-esterified fatty acids (NEFA) and (D) TG concentrations in plasma are shown as mean ± SD, n=5. Independent Student’s t -test.
    Elisa Kit, supplied by ALPCO, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Epiregulin induces leptin secretion and energy expenditure in high-fat diet-fed mice"

    Article Title: Epiregulin induces leptin secretion and energy expenditure in high-fat diet-fed mice

    Journal: The Journal of endocrinology

    doi: 10.1530/JOE-18-0289

    EREG stimulates leptin secretion and lipolysis in mice with diet-induced obesity (DIO). (A) DIO WT male mice (n=7/ group) were injected with 100μL PBS (Veh) with and without EREG (1.5ng/g body weight or 20ng/per iAb depot) into both epididymal iAb fat pads every other day for 2 weeks. Total mRNA was isolated from one whole iAb fad pad. Expression of Lep was measured using NanoString assay. Normalized data represent mean±SD, n=5; Mann-Whitney U test. (B) Expression of LEP protein levels were measured in plasma in the same mice group by ELISA. Data (mean±SD, n=5/group). Independent Student’s t -test. (C) Non-esterified fatty acids (NEFA) and (D) TG concentrations in plasma are shown as mean ± SD, n=5. Independent Student’s t -test.
    Figure Legend Snippet: EREG stimulates leptin secretion and lipolysis in mice with diet-induced obesity (DIO). (A) DIO WT male mice (n=7/ group) were injected with 100μL PBS (Veh) with and without EREG (1.5ng/g body weight or 20ng/per iAb depot) into both epididymal iAb fat pads every other day for 2 weeks. Total mRNA was isolated from one whole iAb fad pad. Expression of Lep was measured using NanoString assay. Normalized data represent mean±SD, n=5; Mann-Whitney U test. (B) Expression of LEP protein levels were measured in plasma in the same mice group by ELISA. Data (mean±SD, n=5/group). Independent Student’s t -test. (C) Non-esterified fatty acids (NEFA) and (D) TG concentrations in plasma are shown as mean ± SD, n=5. Independent Student’s t -test.

    Techniques Used: Mouse Assay, Injection, Isolation, Expressing, MANN-WHITNEY, Enzyme-linked Immunosorbent Assay

    EREG/EGFR/MAPK axes induced leptin (LEP) secretion from adipose tissues ex vivo. Five iAb epididymal fat explants were excised from WT mice and stimulated. After 2h of stimulation LEP levels were measured in the media by ELISA. This experiment was repeated in three different WT mice. (A) Concentration-dependent secretion of LEP in the media from iAb fat explants stimulated with different concentrations of EREG. Data (mean±SD, n=5) are shown as percent to control (Veh, 100 %). Kruskal Wallis test. (B, C) LEP concentrations in the iAb fat lysates (B) and media (C) released from iAb fat before (triangles) and after (circles) stimulation with EREG (black bar, 50 ng/ml). Dashed and solid lines show mean value (n=3 mice) before and after stimulation. (D) Expression levels of Egfr and Erbb4 in epididymal iAb fat of lean WT mice (mean±SD, n=5) were measured by RT-PCR and normalized by TBP. (E) LEP concentrations in the iAb fat stimulated with EREG (black bar, 50 ng/ml) in the presence and absence of inhibitors of EGFR (10 μM) and MAPK (10 μM). Data (mean±SD, n=5) are shown as percent to control (Veh, white bar, 100%, dashed line). Between subject ANOVA with post-hoc Fisher LSD group comparison (α=0.05). (F) LEP in the media released following stimulation of mouse explants with and without EREG (50 ng/mL) in the presence and absence of inhibitors of HSL (10 μM), PPARα (10 μM), and PI3K (100 nM). Data (mean±SD, n=5) are shown as percent of control (Veh, 100%, dashed line). Between subject ANOVA. ns : not significant.
    Figure Legend Snippet: EREG/EGFR/MAPK axes induced leptin (LEP) secretion from adipose tissues ex vivo. Five iAb epididymal fat explants were excised from WT mice and stimulated. After 2h of stimulation LEP levels were measured in the media by ELISA. This experiment was repeated in three different WT mice. (A) Concentration-dependent secretion of LEP in the media from iAb fat explants stimulated with different concentrations of EREG. Data (mean±SD, n=5) are shown as percent to control (Veh, 100 %). Kruskal Wallis test. (B, C) LEP concentrations in the iAb fat lysates (B) and media (C) released from iAb fat before (triangles) and after (circles) stimulation with EREG (black bar, 50 ng/ml). Dashed and solid lines show mean value (n=3 mice) before and after stimulation. (D) Expression levels of Egfr and Erbb4 in epididymal iAb fat of lean WT mice (mean±SD, n=5) were measured by RT-PCR and normalized by TBP. (E) LEP concentrations in the iAb fat stimulated with EREG (black bar, 50 ng/ml) in the presence and absence of inhibitors of EGFR (10 μM) and MAPK (10 μM). Data (mean±SD, n=5) are shown as percent to control (Veh, white bar, 100%, dashed line). Between subject ANOVA with post-hoc Fisher LSD group comparison (α=0.05). (F) LEP in the media released following stimulation of mouse explants with and without EREG (50 ng/mL) in the presence and absence of inhibitors of HSL (10 μM), PPARα (10 μM), and PI3K (100 nM). Data (mean±SD, n=5) are shown as percent of control (Veh, 100%, dashed line). Between subject ANOVA. ns : not significant.

    Techniques Used: Ex Vivo, Mouse Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Expressing, Reverse Transcription Polymerase Chain Reaction

    2) Product Images from "DC260126: A Small-Molecule Antagonist of GPR40 that Protects against Pancreatic ?-Cells Dysfunction in db/db Mice"

    Article Title: DC260126: A Small-Molecule Antagonist of GPR40 that Protects against Pancreatic ?-Cells Dysfunction in db/db Mice

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0066744

    The expression of insulin/proinsulin and P/I ratio analysis in db/db mice. (A) Double immunofluorescence of insulin (green fluorescence) and proinsulin (red fluorescence) from pancreas in different groups, the merged figures were shown in right. The scale bar = 100 µm, and referred to all panels. (B) The blood proinsulin concentrations were measured by ELISA kit, and proinsulin-to-insulin ratio were shown in the graph (C). All data are mean ± SE and * indicated significant differences (* p
    Figure Legend Snippet: The expression of insulin/proinsulin and P/I ratio analysis in db/db mice. (A) Double immunofluorescence of insulin (green fluorescence) and proinsulin (red fluorescence) from pancreas in different groups, the merged figures were shown in right. The scale bar = 100 µm, and referred to all panels. (B) The blood proinsulin concentrations were measured by ELISA kit, and proinsulin-to-insulin ratio were shown in the graph (C). All data are mean ± SE and * indicated significant differences (* p

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

    3) Product Images from "An expanded myeloid derived suppressor cell population does not play a role in gammaherpesvirus-exacerbated breast cancer metastases"

    Article Title: An expanded myeloid derived suppressor cell population does not play a role in gammaherpesvirus-exacerbated breast cancer metastases

    Journal: Infectious Agents and Cancer

    doi: 10.1186/1750-9378-7-22

    ELISA quantification of S100A8 and S100A9 in sera of mock treated or HV-68 infected mice with metastatic breast cancer. Groups of mice were mock treated or HV-68 infected. Six months post-infection, mice were injected with syngeneic 4 T1 mammary tumor cells into the mammary fat pad. At day 44 following tumor cell transplantation, groups of mice were euthanized and sera isolated from individual animals. Each serum sample was assayed for the presence of S100A8 and S100A9 using an ELISA capable of recognizing either protein. Results are presented as means (N = 6) ± standard errors
    Figure Legend Snippet: ELISA quantification of S100A8 and S100A9 in sera of mock treated or HV-68 infected mice with metastatic breast cancer. Groups of mice were mock treated or HV-68 infected. Six months post-infection, mice were injected with syngeneic 4 T1 mammary tumor cells into the mammary fat pad. At day 44 following tumor cell transplantation, groups of mice were euthanized and sera isolated from individual animals. Each serum sample was assayed for the presence of S100A8 and S100A9 using an ELISA capable of recognizing either protein. Results are presented as means (N = 6) ± standard errors

    Techniques Used: Enzyme-linked Immunosorbent Assay, Infection, Mouse Assay, Injection, Transplantation Assay, Isolation

    Related Articles

    Enzyme-linked Immunosorbent Assay:

    Article Title: Cathepsin K deficiency ameliorates systemic lupus erythematosus-like manifestations in Faslpr mice
    Article Snippet: .. The ELISA kits used in this study include: mouse CatK (ALPCO, Salem, NH), mouse IL2 DuoSet (DY402, R & D systems, Minneapolis, MN) and mouse IFN-γ ELISA (555138, BD Biosciences). .. The following antibodies were used for FACS analysis of splenocyte preparation: FcR-blocking antibody anti-CD16/32 (eBioscience), anti-CD4–Alexa Fluor 488, anti-CD25–PE, anti-mFoxp3–Alexa Fluor 647, anti-mouse–PE and all isotype controls (all from BD Biosciences).

    Article Title: Disruption of Nrf2, a Key Inducer of Antioxidant Defenses, Attenuates ApoE-Mediated Atherosclerosis in Mice
    Article Snippet: Glucose was measured in serum using the ACE glucose reagent (Alfa Wassermann, West Caldwell, NJ) and analyzed using the ACE chemistry analyzer. .. Serum oxLDL and Lipid Peroxidation Measurements MDA-modified oxLDL was measured using the oxidized LDL ELISA kit (ALPCO Diagnostics, Salem, NH), as described by the manufacturer. .. For lipid peroxidation measurements, liver tissues were isolated and homogenized in 10 µL/mg ice-cold PBS containing 5 mM butylated hydroxytoluene to inhibit ex vivo oxidation.

    Article Title: TGFβ1 alters androgenic metabolites and hydroxysteroid dehydrogenase enzyme expression in human prostate reactive stromal primary cells: Is steroid metabolism altered by prostate reactive stromal microenvironment?
    Article Snippet: The conditioned culture media was collected for ELISA analysis of DHEA metabolites and the cells were subject to RNA isolation. .. DHEA metabolites were detected by using ELISA kits for free testosterone (cat # 11-FTSHU-E01-ALPCO, Salem, NH) and androstenedione (cat # DSL-10-3800-Beckman Coulter DSLabs, Webster, TX) according to the manufacturer’s instructions. ..

    Article Title: ZBTB20 is required for anterior pituitary development and lactotrope specification
    Article Snippet: RNA interference analysis After wash with PBS twice, 5 × 105 GH3 or MMQ cells were resuspended in BTXpress High Performance Electroporation solution (Harvard Apparatus, MA, USA) containing 2.5 μmol of control or Zbtb20 siRNA (Dhmarcon, PA, USA), and subjected to electroporation in a 2-mm cuvette using a BTX ECM830 electroporator , with the setting of one pulse of 170 volts for 15 ms. Then cells were plated in normal culture media. .. After culture for another 48 h at 37 °C, the culture supernatants were collected for PRL measure with ELISA kit (Alpco), and the cells were subjected to mRNA and protein expression analyses. .. Chromatin immunoprecipitation analysis Fragmented chromatin from GH3 or MMQ cells was incubated overnight with anti-ZBTB20 antibody 9A10 (home-made, 0.5 μg per 106 cells), anti-acetylated histone 3 (Millipore, 0.5 μg per 106 cells) as positive control, or isotype IgG (Upstate, 0.5 μg per 106 cells) as negative control, and followed by incubation with Dynabead-conjugated Protein G (Invitrogen).

    Article Title: Growth Hormone Upregulates Mediators of Melanoma Drug Efflux and Epithelial-to-Mesenchymal Transition In Vitro and In Vivo
    Article Snippet: For tumor GH and IGF-1 measurements, lysed tumor proteins were collected and quantified by Bradford assay. .. GH and IGF-1 were quantified by ELISA kits (#22-GHOMS-E01; #22-IG1MS-E01, ALPCO) and normalized to total protein concentrations. ..

    Article Title: Absence of Functional Leptin Receptor Isoforms in the POUND (Leprdb/lb) Mouse Is Associated with Muscle Atrophy and Altered Myoblast Proliferation and Differentiation
    Article Snippet: IGF-1 ELISA kits were purchased from R & D Systems. .. Myostatin ELISA kits were purchased from Alpco diagnostic and performed according to manufacturer's protocol as we have described previously (33). ..

    Article Title: Dual and opposing roles of the unfolded protein response regulated by IRE1? and XBP1 in proinsulin processing and insulin secretion
    Article Snippet: After removing insoluble materials, insulin levels in pancreas extracts were measured by ELISA as above. .. Serum proinsulin levels were measured using an ELISA kit (ALPCO Diagnostics). .. For immunohistochemistry, we used guinea pig anti-insulin (4011-01; Linco), rabbit anti-glucagon (AB932; Chemicon), mouse anti-CPE (610758; BD Biosciences), rabbit anti-Glut2 (07-1402; Millipore), and mouse anti-BrdU (Bu20a; Biolegend).

    Article Title: Astragaloside IV alleviates placental oxidative stress and inflammation in GDM mice
    Article Snippet: Superoxide dismutase (SOD) activity was detected using a xanthine oxidase technique kit (ShanghaiSolarbio Bioscience and Technology Co., Shanghai, China). .. ELISAInterleukin-6 (IL-6), IL-1β, tumor necrosis factor-α (TNF-α) and serum insulin levels were measured using ELISA kits according to the manufacturer’s instructions, respectively (AlPCO, France). .. Western blot Protein was extracted from lysed placenta in radioimmunoprecipitation lysis buffer on GD 20.

    Multiple Displacement Amplification:

    Article Title: Disruption of Nrf2, a Key Inducer of Antioxidant Defenses, Attenuates ApoE-Mediated Atherosclerosis in Mice
    Article Snippet: Glucose was measured in serum using the ACE glucose reagent (Alfa Wassermann, West Caldwell, NJ) and analyzed using the ACE chemistry analyzer. .. Serum oxLDL and Lipid Peroxidation Measurements MDA-modified oxLDL was measured using the oxidized LDL ELISA kit (ALPCO Diagnostics, Salem, NH), as described by the manufacturer. .. For lipid peroxidation measurements, liver tissues were isolated and homogenized in 10 µL/mg ice-cold PBS containing 5 mM butylated hydroxytoluene to inhibit ex vivo oxidation.

    Expressing:

    Article Title: ZBTB20 is required for anterior pituitary development and lactotrope specification
    Article Snippet: RNA interference analysis After wash with PBS twice, 5 × 105 GH3 or MMQ cells were resuspended in BTXpress High Performance Electroporation solution (Harvard Apparatus, MA, USA) containing 2.5 μmol of control or Zbtb20 siRNA (Dhmarcon, PA, USA), and subjected to electroporation in a 2-mm cuvette using a BTX ECM830 electroporator , with the setting of one pulse of 170 volts for 15 ms. Then cells were plated in normal culture media. .. After culture for another 48 h at 37 °C, the culture supernatants were collected for PRL measure with ELISA kit (Alpco), and the cells were subjected to mRNA and protein expression analyses. .. Chromatin immunoprecipitation analysis Fragmented chromatin from GH3 or MMQ cells was incubated overnight with anti-ZBTB20 antibody 9A10 (home-made, 0.5 μg per 106 cells), anti-acetylated histone 3 (Millipore, 0.5 μg per 106 cells) as positive control, or isotype IgG (Upstate, 0.5 μg per 106 cells) as negative control, and followed by incubation with Dynabead-conjugated Protein G (Invitrogen).

    Diagnostic Assay:

    Article Title: Absence of Functional Leptin Receptor Isoforms in the POUND (Leprdb/lb) Mouse Is Associated with Muscle Atrophy and Altered Myoblast Proliferation and Differentiation
    Article Snippet: IGF-1 ELISA kits were purchased from R & D Systems. .. Myostatin ELISA kits were purchased from Alpco diagnostic and performed according to manufacturer's protocol as we have described previously (33). ..

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    ALPCO elisa kit
    Impaired insulin secretion and <t>proinsulin</t> maturation in XBP1-deficient β-cells. ( A ) Glucose-stimulated insulin secretion assays. Serum insulin levels were measured in control heterozygous Xbp1 f/+ ;RIP-cre and Xbp1 f/f ;RIP-cre mice at indicated time points after a bolus glucose injection. Each line represents an individual mouse. ( B ) Min6 cells stably expressing shRNAs targeting control luciferase or XBP1 mRNAs were tested for XBP1s expression by Western blot. ( C ) Cells were pretreated with 2.5 mM glucose for 2 h and then cultured in 5 mM or 25 mM glucose media for 2 h. Culture media were collected to measure insulin content by <t>ELISA.</t> ( D ) Serum proinsulin levels relative to total insulin in WT and Xbp1 f/f ;RIP-cre mice were determined. Data from both males and females were combined, because sex difference was not significant. The graphs display proinsulin:insulin ratio ( Left ) or proinsulin and insulin concentrations (pmol per liter) ( Right ). Each dot represents an individual mouse (4- to 6-mo-old). ( E ) Min6 cells expressing control or XBP1 shRNA were pulse-labeled for 30 min with [ 35 S]Met/Cys and then cultured in chase media for the indicated time. Cells and culture supernatants were harvested for immunoprecipitation of radiolabeled insulin and proinsulin, which were revealed by SDS/PAGE followed by autoradiography.
    Elisa Kit, supplied by ALPCO, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elisa kit/product/ALPCO
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    elisa kit - by Bioz Stars, 2021-03
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    96
    ALPCO insulin elisa kits
    STF improves Akita islet β-cell function. A . Glucose-stimulated insulin secretion of 20 islets isolated from Akita mice treated with STF or vehicle and incubated with 2.5 mM and 16.7 mM glucose. Secreted insulin was measured by <t>ELISA.</t> The data was presented as fold change and normalized with total protein concentration, with the amount of insulin secreted in response to 2.5 mM glucose from vehicle-treated group set to 1.0. B-E . mRNA levels for indicated genes were analyzed in islets isolated from Akita mice treated with STF or vehicle by qRT-PCR. The results are expressed as fold change and are representative of 3 independent experiments. F . <t>Proinsulin</t> content measurement by ELISA as detailed in Methods and Materials. G-H. Proinsulin content and secretion measurement. 20 islets isolated from Akita mice treated with STF or vehicle were incubated with 2.5 mM and 16.7 mM glucose. Secreted proinsulin was measured by ELISA. Proinsulin content measurement by ELISA as detailed in Methods and Materials. The data was presented as fold change and normalized with total protein concentration, with the amount of proinsulin in response to 2.5 mM glucose from vehicle-treated group set to 1.0. I-K . Ultrastructure of β-cells in islets isolated from Akita treated with STF by transmission electron microscopy. Images at the top panel and the bottom panel were taken at 3,000x and 10,000x, respectively. Arrows point to dark mature insulin granules.
    Insulin Elisa Kits, supplied by ALPCO, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    insulin elisa kits - by Bioz Stars, 2021-03
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    Impaired insulin secretion and proinsulin maturation in XBP1-deficient β-cells. ( A ) Glucose-stimulated insulin secretion assays. Serum insulin levels were measured in control heterozygous Xbp1 f/+ ;RIP-cre and Xbp1 f/f ;RIP-cre mice at indicated time points after a bolus glucose injection. Each line represents an individual mouse. ( B ) Min6 cells stably expressing shRNAs targeting control luciferase or XBP1 mRNAs were tested for XBP1s expression by Western blot. ( C ) Cells were pretreated with 2.5 mM glucose for 2 h and then cultured in 5 mM or 25 mM glucose media for 2 h. Culture media were collected to measure insulin content by ELISA. ( D ) Serum proinsulin levels relative to total insulin in WT and Xbp1 f/f ;RIP-cre mice were determined. Data from both males and females were combined, because sex difference was not significant. The graphs display proinsulin:insulin ratio ( Left ) or proinsulin and insulin concentrations (pmol per liter) ( Right ). Each dot represents an individual mouse (4- to 6-mo-old). ( E ) Min6 cells expressing control or XBP1 shRNA were pulse-labeled for 30 min with [ 35 S]Met/Cys and then cultured in chase media for the indicated time. Cells and culture supernatants were harvested for immunoprecipitation of radiolabeled insulin and proinsulin, which were revealed by SDS/PAGE followed by autoradiography.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Dual and opposing roles of the unfolded protein response regulated by IRE1? and XBP1 in proinsulin processing and insulin secretion

    doi: 10.1073/pnas.1105564108

    Figure Lengend Snippet: Impaired insulin secretion and proinsulin maturation in XBP1-deficient β-cells. ( A ) Glucose-stimulated insulin secretion assays. Serum insulin levels were measured in control heterozygous Xbp1 f/+ ;RIP-cre and Xbp1 f/f ;RIP-cre mice at indicated time points after a bolus glucose injection. Each line represents an individual mouse. ( B ) Min6 cells stably expressing shRNAs targeting control luciferase or XBP1 mRNAs were tested for XBP1s expression by Western blot. ( C ) Cells were pretreated with 2.5 mM glucose for 2 h and then cultured in 5 mM or 25 mM glucose media for 2 h. Culture media were collected to measure insulin content by ELISA. ( D ) Serum proinsulin levels relative to total insulin in WT and Xbp1 f/f ;RIP-cre mice were determined. Data from both males and females were combined, because sex difference was not significant. The graphs display proinsulin:insulin ratio ( Left ) or proinsulin and insulin concentrations (pmol per liter) ( Right ). Each dot represents an individual mouse (4- to 6-mo-old). ( E ) Min6 cells expressing control or XBP1 shRNA were pulse-labeled for 30 min with [ 35 S]Met/Cys and then cultured in chase media for the indicated time. Cells and culture supernatants were harvested for immunoprecipitation of radiolabeled insulin and proinsulin, which were revealed by SDS/PAGE followed by autoradiography.

    Article Snippet: Serum proinsulin levels were measured using an ELISA kit (ALPCO Diagnostics).

    Techniques: Mouse Assay, Injection, Stable Transfection, Expressing, Luciferase, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay, shRNA, Labeling, Immunoprecipitation, SDS Page, Autoradiography

    Subcutaneous B16-F10 mouse melanoma growth in bGH and GHRKO syngeneic mice. ( A ) Subcutaneous B16-F10 tumor growth in male bGH (n = 5) vs. WT mice (n = 6). ( B ) Subcutaneous B16-F10 tumor growth in female bGH (n = 7) and WT mice (n = 6). ( C ) Subcutaneous B16-F10 tumor growth in male GHRKO (n = 6) vs. WT (n = 8) mice. ( D ) Subcutaneous B16-F10 tumor growth in female GHRKO (n = 4) and WT mice (n = 5). Tumor sizes were analyzed by repeated measures (SPSS). ( E ) Weight of subcutaneous B16-F10 tumors from bGH vs. WT mice at dissection. ( F ) Weight of subcutaneous B16-F10 tumors from GHRKO vs. WT mice at dissection. ( G ) GH levels were measured in protein lysates isolated from tumors of bGH and WT mice using ELISA and normalized to total protein concentrations (n = 4). ( H ) Similar GH measurements were performed in protein lysates isolated from tumors of GHRKO and WT mice (males n = 3, females n = 4). ( I ) IGF-1 levels were measured in protein lysates isolated from tumors of bGH and WT mice using ELISA and normalized to total protein concentrations (n = 4). ( J ) Similar IGF-1 measurements were performed in protein lysates isolated from tumors of GHRKO and WT mice (males n = 3, females n = 4). ( K ) Representative images of western blot analysis of phosphorylation (p) and total (t) levels of STAT1, STAT3 and STAT5 in protein lysates isolated from tumors of bGH and WT mice. Densitometry analysis was performed and normalized against β-Actin. The relative expression levels (fold change relative to WT) are labeled under each band. The ratio of phosphorylated vs. total protein levels in tumors from bGH and WT mice are presented in bar graphs (n = 4). ( L ) Representative images of western blot analysis of phosphorylation and total levels of STAT1, STAT3, and STAT5 in protein lysates isolated from tumors of GHRKO and WT mice. (males n = 3, females n = 4). Data are presented as mean ± standard errors (*, p

    Journal: Cancers

    Article Title: Growth Hormone Upregulates Mediators of Melanoma Drug Efflux and Epithelial-to-Mesenchymal Transition In Vitro and In Vivo

    doi: 10.3390/cancers12123640

    Figure Lengend Snippet: Subcutaneous B16-F10 mouse melanoma growth in bGH and GHRKO syngeneic mice. ( A ) Subcutaneous B16-F10 tumor growth in male bGH (n = 5) vs. WT mice (n = 6). ( B ) Subcutaneous B16-F10 tumor growth in female bGH (n = 7) and WT mice (n = 6). ( C ) Subcutaneous B16-F10 tumor growth in male GHRKO (n = 6) vs. WT (n = 8) mice. ( D ) Subcutaneous B16-F10 tumor growth in female GHRKO (n = 4) and WT mice (n = 5). Tumor sizes were analyzed by repeated measures (SPSS). ( E ) Weight of subcutaneous B16-F10 tumors from bGH vs. WT mice at dissection. ( F ) Weight of subcutaneous B16-F10 tumors from GHRKO vs. WT mice at dissection. ( G ) GH levels were measured in protein lysates isolated from tumors of bGH and WT mice using ELISA and normalized to total protein concentrations (n = 4). ( H ) Similar GH measurements were performed in protein lysates isolated from tumors of GHRKO and WT mice (males n = 3, females n = 4). ( I ) IGF-1 levels were measured in protein lysates isolated from tumors of bGH and WT mice using ELISA and normalized to total protein concentrations (n = 4). ( J ) Similar IGF-1 measurements were performed in protein lysates isolated from tumors of GHRKO and WT mice (males n = 3, females n = 4). ( K ) Representative images of western blot analysis of phosphorylation (p) and total (t) levels of STAT1, STAT3 and STAT5 in protein lysates isolated from tumors of bGH and WT mice. Densitometry analysis was performed and normalized against β-Actin. The relative expression levels (fold change relative to WT) are labeled under each band. The ratio of phosphorylated vs. total protein levels in tumors from bGH and WT mice are presented in bar graphs (n = 4). ( L ) Representative images of western blot analysis of phosphorylation and total levels of STAT1, STAT3, and STAT5 in protein lysates isolated from tumors of GHRKO and WT mice. (males n = 3, females n = 4). Data are presented as mean ± standard errors (*, p

    Article Snippet: GH and IGF-1 were quantified by ELISA kits (#22-GHOMS-E01; #22-IG1MS-E01, ALPCO) and normalized to total protein concentrations.

    Techniques: Mouse Assay, Dissection, Isolation, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Labeling

    Astragaloside IV reduced placental inflammation in the late stage of pregnancy in GDM mice. ELISA was used to analyze the levels of IL-6 (A), IL-1β (B) and TNF-α (C) in placenta on GD 20. qRT-PCR was used to analyzed the mRNA levels of IL-6 (D), IL-1β (E) and TNF-α (F) in placenta on GD 20. GAPDH was set as a loading control and the relative expressions were normalized to wild type. Data were presented as means ± s.e.m. n = 6 for each group. ## P

    Journal: Endocrine Connections

    Article Title: Astragaloside IV alleviates placental oxidative stress and inflammation in GDM mice

    doi: 10.1530/EC-20-0295

    Figure Lengend Snippet: Astragaloside IV reduced placental inflammation in the late stage of pregnancy in GDM mice. ELISA was used to analyze the levels of IL-6 (A), IL-1β (B) and TNF-α (C) in placenta on GD 20. qRT-PCR was used to analyzed the mRNA levels of IL-6 (D), IL-1β (E) and TNF-α (F) in placenta on GD 20. GAPDH was set as a loading control and the relative expressions were normalized to wild type. Data were presented as means ± s.e.m. n = 6 for each group. ## P

    Article Snippet: ELISAInterleukin-6 (IL-6), IL-1β, tumor necrosis factor-α (TNF-α) and serum insulin levels were measured using ELISA kits according to the manufacturer’s instructions, respectively (AlPCO, France).

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

    STF improves Akita islet β-cell function. A . Glucose-stimulated insulin secretion of 20 islets isolated from Akita mice treated with STF or vehicle and incubated with 2.5 mM and 16.7 mM glucose. Secreted insulin was measured by ELISA. The data was presented as fold change and normalized with total protein concentration, with the amount of insulin secreted in response to 2.5 mM glucose from vehicle-treated group set to 1.0. B-E . mRNA levels for indicated genes were analyzed in islets isolated from Akita mice treated with STF or vehicle by qRT-PCR. The results are expressed as fold change and are representative of 3 independent experiments. F . Proinsulin content measurement by ELISA as detailed in Methods and Materials. G-H. Proinsulin content and secretion measurement. 20 islets isolated from Akita mice treated with STF or vehicle were incubated with 2.5 mM and 16.7 mM glucose. Secreted proinsulin was measured by ELISA. Proinsulin content measurement by ELISA as detailed in Methods and Materials. The data was presented as fold change and normalized with total protein concentration, with the amount of proinsulin in response to 2.5 mM glucose from vehicle-treated group set to 1.0. I-K . Ultrastructure of β-cells in islets isolated from Akita treated with STF by transmission electron microscopy. Images at the top panel and the bottom panel were taken at 3,000x and 10,000x, respectively. Arrows point to dark mature insulin granules.

    Journal: bioRxiv

    Article Title: Inhibition of inositol-requiring enzyme 1α RNase activity protects pancreatic beta cell and improves diabetic condition in insulin mutation-induced diabetes

    doi: 10.1101/2020.08.30.274381

    Figure Lengend Snippet: STF improves Akita islet β-cell function. A . Glucose-stimulated insulin secretion of 20 islets isolated from Akita mice treated with STF or vehicle and incubated with 2.5 mM and 16.7 mM glucose. Secreted insulin was measured by ELISA. The data was presented as fold change and normalized with total protein concentration, with the amount of insulin secreted in response to 2.5 mM glucose from vehicle-treated group set to 1.0. B-E . mRNA levels for indicated genes were analyzed in islets isolated from Akita mice treated with STF or vehicle by qRT-PCR. The results are expressed as fold change and are representative of 3 independent experiments. F . Proinsulin content measurement by ELISA as detailed in Methods and Materials. G-H. Proinsulin content and secretion measurement. 20 islets isolated from Akita mice treated with STF or vehicle were incubated with 2.5 mM and 16.7 mM glucose. Secreted proinsulin was measured by ELISA. Proinsulin content measurement by ELISA as detailed in Methods and Materials. The data was presented as fold change and normalized with total protein concentration, with the amount of proinsulin in response to 2.5 mM glucose from vehicle-treated group set to 1.0. I-K . Ultrastructure of β-cells in islets isolated from Akita treated with STF by transmission electron microscopy. Images at the top panel and the bottom panel were taken at 3,000x and 10,000x, respectively. Arrows point to dark mature insulin granules.

    Article Snippet: The secreted insulin and proinsulin from supernatant were measured with insulin ELISA kits (ALPCO, Salem, NH) and Rat/Mouse Proinsulin kit (Mercodia), respectively. islets were lysed with RIPA buffer (50 mM Tris HCl pH 7.4, 1% NP-40, 0.25% sodium deoxycholate, 150 mM NaCl), and total cellular protein was determined with a Bradford protein assay.

    Techniques: Cell Function Assay, Isolation, Mouse Assay, Incubation, Enzyme-linked Immunosorbent Assay, Protein Concentration, Quantitative RT-PCR, Transmission Assay, Electron Microscopy