anti goat  (Abcam)

 
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    Name:
    Goat F ab 2 Anti Mouse IgG IgM IgA H L HRP
    Description:

    Catalog Number:
    ab6006
    Price:
    None
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    Structured Review

    Abcam anti goat

    https://www.bioz.com/result/anti goat/product/Abcam
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    anti goat - by Bioz Stars, 2020-09
    99/100 stars

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    Related Articles

    Western Blot:

    Article Title: In planta expression of nanobody-based designer chicken antibodies targeting Campylobacter
    Article Snippet: .. Subsequently, the western blot was developed using goat anti-IgA (1/5000) or goat anti-IgY (1/1250) conjugated to HRP (Abcam). .. Immunofluorescence microscopy C . jejuni bacteria were grown microaerobically on NB2 agar plates.

    Article Title: The Tick Protein Sialostatin L2 Binds to Annexin A2 and Inhibits NLRC4-Mediated Inflammasome Activation
    Article Snippet: .. Western blot antibodies for caspase-1 (1:1,000 [catalog number 06-503 or 06-503-I; Millipore], 1:2,000 [catalog number 4175, cell line 4B4.2.1; Genentech], or 1:1,000 [catalog number AG-20B-0042; AdiPoGen International]), NLRC4 (1:1,000) (catalog number 06-1125; Millipore), IL-1β (1:1,000) (catalog numbers AF401-NA [R & D Systems] and 12426S [Cell Signaling]), IL-18 (1:1,000) (catalog number JM-5180-100; MBL), β-actin (1:1,000) (catalog number A2103; Sigma), annexin A2 (1:1,000) (catalog numbers 60051-1-1g [Protein Tech] and PA1-9006 [Thermo Scientific]), anti-mouse horseradish peroxidase (HRP), anti-goat HRP, anti-rabbit HRP (1:5,000) (catalog numbers ab97046, ab97110, and ab97051, respectively; Abcam), and anti-rat (1:5,000) (catalog numbers ab97057 [Abcam] and sc-2006 [Santa Cruz Biotechnology]) were used. .. The enhanced chemiluminescence (ECL) Western blotting substrate and Super Signal West Pico chemiluminescent substrate were used (Thermo Scientific).

    Immunohistochemistry:

    Article Title: Markers of fibrosis and epithelial to mesenchymal transition demonstrate field cancerization in histologically normal tissue adjacent to breast tumors
    Article Snippet: .. For IHC, secondary antibodies used were goat anti rabbit-HRP and goat antimouse-HRP (Abcam). .. HRP activity was visualized using the Liquid DAB Plus Substrate Kit (Zymed) according to manufacturer’s instructions.

    Incubation:

    Article Title: Microparticles as Potential Mediators of High Glucose-Induced Renal Cell Injury
    Article Snippet: .. Subsequently, membranes were incubated with horseradish peroxidase (HRP)-conjugated goat antimouse IgG or goat antirabbit IgG secondary antibodies (Abcam) for 1 h at room temperature. .. The immunoreactivity was visualized with Optiblot ECL detection kit (Abcam) using FluorChemTM M imaging system (Protein Simple, San Jose, CA, USA) and analyzed using AlphaView software (Protein Simple).

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  • 93
    Abcam goat anti mouse igg
    Internalization of WB‐6 into living human monocytic leukemia cell line THP‐1 cells and human umbilical vein endothelial cells (HUVECs). (a) THP‐1 cells were incubated with 25 µg/ml WB‐6, isotype‐matched control (IC) or 5 µg/ml 2C10 for 2 h. After washing, fixation, permeabilization and blocking, internalized immunoglobulin (Ig)G was detected using <t>Alexa</t> Fluor 488‐labeled goat anti‐mouse <t>IgG</t> (green). (b) THP‐1 cells were incubated with (center column) or without (left column) 25 µg/ml WB‐6 for 2 h or 1 µM staurosporine for 6 h (right column), and expression of phosphatidylserine was tested using annexin V‐biotin and fluorescein isothiocyanate (FITC)‐streptavidin (green). (c) THP‐1 cells were incubated with 25 µg/ml WB‐6 for 2 h, and Alexa Fluor 488‐labeled goat anti‐mouse IgG (green) was added after (left) or before (right) fixation and permeabilization. (d) Internalization of WB‐6 into HUVECs was tested according to the same protocol as (a). In all experiments, the nuclei were stained with Hoechst 33342 (blue). A representative of three independent experiments with similar results is shown.
    Goat Anti Mouse Igg, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 86 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti mouse igg/product/Abcam
    Average 93 stars, based on 86 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse igg - by Bioz Stars, 2020-09
    93/100 stars
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    91
    Abcam goat anti sec61a1 antibody
    Mutant Sec61α1 sensitizes MIN6 cells to the effects of ER stress. A : MIN6 cells were transfected with either ERSE (left) or UPRE (right) firefly luciferase plasmid, the indicated Sec61 expression plasmid, and a CMV Renilla luciferase reporter plasmid treated with 1 μmol/l thapsigargin and assayed for reporter activation after 16 h using a luminometer. Assays were performed in triplicate and normalized to CMV Renilla activity. Values represent the means ± SD. B : MIN6 cells were transfected with the indicated Sec61 expression plasmid and then treated with 1 μmol/l thapsigargin for 4 h. Cells were lysed and Western blotting was performed with the indicated antibodies. Quantification of the Western signals is shown below the blots. C : Pulse-chase analysis of insulin processing in MIN6 cells transfected with the indicted expression plasmids. Cells were labeled for 20 min with 50 μCi of 35 S-labeled Met and chased for the indicated time points. The upper band corresponds to proinsulin, whereas the lower band corresponds to the fully processed form. D: <t>Sec61a1</t> and Bip gene expression in the MIN6 β-cell line and their upregulation in cells cultured in high-glucose media or in the presence of the ER stress-inducer thapsigargin (Tg). The x -axis shows the time in hours after the initial exposure of the cells to Tg or high glucose. Relative expression levels were determined by comparative Ct analysis. Values represent means ± SD.
    Goat Anti Sec61a1 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti sec61a1 antibody/product/Abcam
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti sec61a1 antibody - by Bioz Stars, 2020-09
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    99
    Abcam anti rabbit igg
    Recombinant ACKR3 and <t>AVPR1A</t> form heteromeric complexes. ( a ) Typical PLA images for the detection of individual receptors and receptor–receptor interactions in HEK293T cells transfected with DNA encoding HA- or FLAG-tagged receptors. Ctrl: cells transfected with pcDNA. Images show merged PLA/DAPI signals acquired from z -stack images ( n = 10; thickness 1 µm, bottom to top). Scale bars, 10 µm. ( b ) Quantification of PLA signals per cell as in ( a ). n = 3 independent experiments with n = 10 images per condition and experiment. ( c ) HEK293T cells expressing HA-AVPR1A and FLAG-ACKR3 were lysed (input), the lysate was immunoprecipitated (IP) with anti-HA, followed by immunoblotting (IB) to detect HA-AVPR1A (left) and FLAG-ACKR3 (right) in the IP samples. IP control: precipitate after incubation of cell lysates with <t>IgG-coupled</t> resin. ( d , e ) Intermolecular BRET assays. Cells were co-transfected with AVPR1A-hRluc plus EYFP (open circles) or ACKR3-EYFP (grey squares) at various acceptor : donor ratios ( d ) and with increasing amounts of AVPR1A-hRluc and ACKR3-EYFP at a constant ratio of 1 : 10 ( e ).
    Anti Rabbit Igg, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti rabbit igg/product/Abcam
    Average 99 stars, based on 50 article reviews
    Price from $9.99 to $1999.99
    anti rabbit igg - by Bioz Stars, 2020-09
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    Image Search Results


    Internalization of WB‐6 into living human monocytic leukemia cell line THP‐1 cells and human umbilical vein endothelial cells (HUVECs). (a) THP‐1 cells were incubated with 25 µg/ml WB‐6, isotype‐matched control (IC) or 5 µg/ml 2C10 for 2 h. After washing, fixation, permeabilization and blocking, internalized immunoglobulin (Ig)G was detected using Alexa Fluor 488‐labeled goat anti‐mouse IgG (green). (b) THP‐1 cells were incubated with (center column) or without (left column) 25 µg/ml WB‐6 for 2 h or 1 µM staurosporine for 6 h (right column), and expression of phosphatidylserine was tested using annexin V‐biotin and fluorescein isothiocyanate (FITC)‐streptavidin (green). (c) THP‐1 cells were incubated with 25 µg/ml WB‐6 for 2 h, and Alexa Fluor 488‐labeled goat anti‐mouse IgG (green) was added after (left) or before (right) fixation and permeabilization. (d) Internalization of WB‐6 into HUVECs was tested according to the same protocol as (a). In all experiments, the nuclei were stained with Hoechst 33342 (blue). A representative of three independent experiments with similar results is shown.

    Journal: Clinical and Experimental Immunology

    Article Title: Anti‐β2‐glycoprotein I antibody with DNA binding activity enters living monocytes via cell surface DNA and induces tissue factor expression

    doi: 10.1111/cei.13229

    Figure Lengend Snippet: Internalization of WB‐6 into living human monocytic leukemia cell line THP‐1 cells and human umbilical vein endothelial cells (HUVECs). (a) THP‐1 cells were incubated with 25 µg/ml WB‐6, isotype‐matched control (IC) or 5 µg/ml 2C10 for 2 h. After washing, fixation, permeabilization and blocking, internalized immunoglobulin (Ig)G was detected using Alexa Fluor 488‐labeled goat anti‐mouse IgG (green). (b) THP‐1 cells were incubated with (center column) or without (left column) 25 µg/ml WB‐6 for 2 h or 1 µM staurosporine for 6 h (right column), and expression of phosphatidylserine was tested using annexin V‐biotin and fluorescein isothiocyanate (FITC)‐streptavidin (green). (c) THP‐1 cells were incubated with 25 µg/ml WB‐6 for 2 h, and Alexa Fluor 488‐labeled goat anti‐mouse IgG (green) was added after (left) or before (right) fixation and permeabilization. (d) Internalization of WB‐6 into HUVECs was tested according to the same protocol as (a). In all experiments, the nuclei were stained with Hoechst 33342 (blue). A representative of three independent experiments with similar results is shown.

    Article Snippet: When we added Alexa Fluor 488‐labeled goat anti‐mouse IgG before fixation and permeabilization, virtually all cells remained unstained, as shown in Fig. c. These results again demonstrate the integrity of the cell membrane, because the second antibody could not enter the cells.

    Techniques: Western Blot, Incubation, Blocking Assay, Labeling, Expressing, Staining

    Internalization of WB‐6 into living peripheral blood mononuclear cells (PBMCs). (a) PBMCs from healthy volunteers were incubated with WB‐6 for 2 h and internalized antibody was detected as described in Fig. 2 . A representative immunofluorescence image showing that WB‐6 entered only a small fraction of PBMCs. (b) In flow cytometric analysis, forward‐ and side‐scatter plot of PBMCs showed two populations of the cells, designated P1 and P2, representing monocytes and lymphocytes, respectively. (c) Most of the cells in the P1 gate were confirmed to be CD14 + . (d–f) Representative histograms showing the ratio of Alexa Fluor 488‐positive cells after 2 h incubation of PBMCs with isotype‐matched IgG in the gate P1 (d), WB‐6 in the gate P1 (e) and WB‐6 in the gate P2 (f). (g) The ratio of Alexa Fluor 488‐positive cells in the gate P1 (Mono) and gate P2 (Lym). Data show a representative of three independent experiments with similar results, and the mean ± standard error of the mean (s.e.m.) of triplicate assay.

    Journal: Clinical and Experimental Immunology

    Article Title: Anti‐β2‐glycoprotein I antibody with DNA binding activity enters living monocytes via cell surface DNA and induces tissue factor expression

    doi: 10.1111/cei.13229

    Figure Lengend Snippet: Internalization of WB‐6 into living peripheral blood mononuclear cells (PBMCs). (a) PBMCs from healthy volunteers were incubated with WB‐6 for 2 h and internalized antibody was detected as described in Fig. 2 . A representative immunofluorescence image showing that WB‐6 entered only a small fraction of PBMCs. (b) In flow cytometric analysis, forward‐ and side‐scatter plot of PBMCs showed two populations of the cells, designated P1 and P2, representing monocytes and lymphocytes, respectively. (c) Most of the cells in the P1 gate were confirmed to be CD14 + . (d–f) Representative histograms showing the ratio of Alexa Fluor 488‐positive cells after 2 h incubation of PBMCs with isotype‐matched IgG in the gate P1 (d), WB‐6 in the gate P1 (e) and WB‐6 in the gate P2 (f). (g) The ratio of Alexa Fluor 488‐positive cells in the gate P1 (Mono) and gate P2 (Lym). Data show a representative of three independent experiments with similar results, and the mean ± standard error of the mean (s.e.m.) of triplicate assay.

    Article Snippet: When we added Alexa Fluor 488‐labeled goat anti‐mouse IgG before fixation and permeabilization, virtually all cells remained unstained, as shown in Fig. c. These results again demonstrate the integrity of the cell membrane, because the second antibody could not enter the cells.

    Techniques: Western Blot, Incubation, Immunofluorescence, Flow Cytometry

    Mutant Sec61α1 sensitizes MIN6 cells to the effects of ER stress. A : MIN6 cells were transfected with either ERSE (left) or UPRE (right) firefly luciferase plasmid, the indicated Sec61 expression plasmid, and a CMV Renilla luciferase reporter plasmid treated with 1 μmol/l thapsigargin and assayed for reporter activation after 16 h using a luminometer. Assays were performed in triplicate and normalized to CMV Renilla activity. Values represent the means ± SD. B : MIN6 cells were transfected with the indicated Sec61 expression plasmid and then treated with 1 μmol/l thapsigargin for 4 h. Cells were lysed and Western blotting was performed with the indicated antibodies. Quantification of the Western signals is shown below the blots. C : Pulse-chase analysis of insulin processing in MIN6 cells transfected with the indicted expression plasmids. Cells were labeled for 20 min with 50 μCi of 35 S-labeled Met and chased for the indicated time points. The upper band corresponds to proinsulin, whereas the lower band corresponds to the fully processed form. D: Sec61a1 and Bip gene expression in the MIN6 β-cell line and their upregulation in cells cultured in high-glucose media or in the presence of the ER stress-inducer thapsigargin (Tg). The x -axis shows the time in hours after the initial exposure of the cells to Tg or high glucose. Relative expression levels were determined by comparative Ct analysis. Values represent means ± SD.

    Journal: Diabetes

    Article Title: A Point Mutation in Sec61?1 Leads to Diabetes and Hepatosteatosis in Mice

    doi: 10.2337/db08-1362

    Figure Lengend Snippet: Mutant Sec61α1 sensitizes MIN6 cells to the effects of ER stress. A : MIN6 cells were transfected with either ERSE (left) or UPRE (right) firefly luciferase plasmid, the indicated Sec61 expression plasmid, and a CMV Renilla luciferase reporter plasmid treated with 1 μmol/l thapsigargin and assayed for reporter activation after 16 h using a luminometer. Assays were performed in triplicate and normalized to CMV Renilla activity. Values represent the means ± SD. B : MIN6 cells were transfected with the indicated Sec61 expression plasmid and then treated with 1 μmol/l thapsigargin for 4 h. Cells were lysed and Western blotting was performed with the indicated antibodies. Quantification of the Western signals is shown below the blots. C : Pulse-chase analysis of insulin processing in MIN6 cells transfected with the indicted expression plasmids. Cells were labeled for 20 min with 50 μCi of 35 S-labeled Met and chased for the indicated time points. The upper band corresponds to proinsulin, whereas the lower band corresponds to the fully processed form. D: Sec61a1 and Bip gene expression in the MIN6 β-cell line and their upregulation in cells cultured in high-glucose media or in the presence of the ER stress-inducer thapsigargin (Tg). The x -axis shows the time in hours after the initial exposure of the cells to Tg or high glucose. Relative expression levels were determined by comparative Ct analysis. Values represent means ± SD.

    Article Snippet: Antibody combinations were as follows: rabbit anti-insulin (sc-9168; Santa Cruz Biotechnology) and AlexaFluor 594–conjugated chicken anti-rabbit antibody; goat anti-glucagon antibody (sc-97780; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated donkey anti-goat antibody; goat anti-SEC61A1 antibody (ab1327; Abcam) and AlexaFluor 488–conjugated donkey anti-goat antibody; anti-immunoglobulin heavy chain–binding protein (BIP) antibody (sc-1050; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated donkey anti-goat antibody; anti-C/EBP homologous protein (CHOP) antibody (sc-7351; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated goat anti-mouse antibody.

    Techniques: Mutagenesis, Transfection, Luciferase, Plasmid Preparation, Expressing, Activation Assay, Activity Assay, Western Blot, Pulse Chase, Labeling, Cell Culture

    Transgenic rescue of Sec61a1 Y344H/Y344H normalizes signs of ER stress in β-cells but not hepatocytes. A : Electron microscopy of β-cells from islets isolated from Sec61a1 +/+ (+/+), Sec61a1 Y344H/Y344H (Y344H/Y344H), and Sec61a1 Y344H/Y344H , Tg[RIP-Sec61] (Y344H/Y344H, RIP-Sec61) mice. Bar, 1 μm. g, secretory granules. B : qPCR on islets from Sec61a1 +/+ (+/+) and Sec61a1 Y344H/Y344H , Tg[RIP-Sec61] (Y344H/Y344H, RIP-Sec61) mice. Total RNA was extracted from isolated islets of age- and sex-matched mice from each genotype ( n = 2). Relative expression levels of the indicated genes were determined by comparative Ct analysis. Values shown are means ± SD. Statistical assessments were made using a two-tailed Student t test assuming unequal variances. * P

    Journal: Diabetes

    Article Title: A Point Mutation in Sec61?1 Leads to Diabetes and Hepatosteatosis in Mice

    doi: 10.2337/db08-1362

    Figure Lengend Snippet: Transgenic rescue of Sec61a1 Y344H/Y344H normalizes signs of ER stress in β-cells but not hepatocytes. A : Electron microscopy of β-cells from islets isolated from Sec61a1 +/+ (+/+), Sec61a1 Y344H/Y344H (Y344H/Y344H), and Sec61a1 Y344H/Y344H , Tg[RIP-Sec61] (Y344H/Y344H, RIP-Sec61) mice. Bar, 1 μm. g, secretory granules. B : qPCR on islets from Sec61a1 +/+ (+/+) and Sec61a1 Y344H/Y344H , Tg[RIP-Sec61] (Y344H/Y344H, RIP-Sec61) mice. Total RNA was extracted from isolated islets of age- and sex-matched mice from each genotype ( n = 2). Relative expression levels of the indicated genes were determined by comparative Ct analysis. Values shown are means ± SD. Statistical assessments were made using a two-tailed Student t test assuming unequal variances. * P

    Article Snippet: Antibody combinations were as follows: rabbit anti-insulin (sc-9168; Santa Cruz Biotechnology) and AlexaFluor 594–conjugated chicken anti-rabbit antibody; goat anti-glucagon antibody (sc-97780; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated donkey anti-goat antibody; goat anti-SEC61A1 antibody (ab1327; Abcam) and AlexaFluor 488–conjugated donkey anti-goat antibody; anti-immunoglobulin heavy chain–binding protein (BIP) antibody (sc-1050; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated donkey anti-goat antibody; anti-C/EBP homologous protein (CHOP) antibody (sc-7351; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated goat anti-mouse antibody.

    Techniques: Transgenic Assay, Electron Microscopy, Isolation, Mouse Assay, Real-time Polymerase Chain Reaction, Expressing, Two Tailed Test

    Sec61a1 Y344H/Y344H mice lose β-cells by apoptosis and exhibit signs of ER stress. A: Islets from neonate, 4-, 7-, and 12-week-old wild-type (+/+) and mutant (Y344H/Y344H) mice fed HFD stained for insulin (red), glucagon (green), and nuclei (blue) (×600 magnification). B: Quantitation of islet composition from 12- to 15-week-old sex-matched mice fed HFD ( n = 3 mice each genotype, 10 islets each). Error bars represent SD. C: Quantitative assessment of TUNEL + β-cells in islets from three wild-type (+/+) and three mutant (Y344H/Y344H) mice aged 4 weeks and fed HFD for 1 week. Data are expressed as the number of TUNEL + cells per islet. Statistical assessment was made using a two-tailed Student t test assuming unequal variances. * P

    Journal: Diabetes

    Article Title: A Point Mutation in Sec61?1 Leads to Diabetes and Hepatosteatosis in Mice

    doi: 10.2337/db08-1362

    Figure Lengend Snippet: Sec61a1 Y344H/Y344H mice lose β-cells by apoptosis and exhibit signs of ER stress. A: Islets from neonate, 4-, 7-, and 12-week-old wild-type (+/+) and mutant (Y344H/Y344H) mice fed HFD stained for insulin (red), glucagon (green), and nuclei (blue) (×600 magnification). B: Quantitation of islet composition from 12- to 15-week-old sex-matched mice fed HFD ( n = 3 mice each genotype, 10 islets each). Error bars represent SD. C: Quantitative assessment of TUNEL + β-cells in islets from three wild-type (+/+) and three mutant (Y344H/Y344H) mice aged 4 weeks and fed HFD for 1 week. Data are expressed as the number of TUNEL + cells per islet. Statistical assessment was made using a two-tailed Student t test assuming unequal variances. * P

    Article Snippet: Antibody combinations were as follows: rabbit anti-insulin (sc-9168; Santa Cruz Biotechnology) and AlexaFluor 594–conjugated chicken anti-rabbit antibody; goat anti-glucagon antibody (sc-97780; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated donkey anti-goat antibody; goat anti-SEC61A1 antibody (ab1327; Abcam) and AlexaFluor 488–conjugated donkey anti-goat antibody; anti-immunoglobulin heavy chain–binding protein (BIP) antibody (sc-1050; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated donkey anti-goat antibody; anti-C/EBP homologous protein (CHOP) antibody (sc-7351; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated goat anti-mouse antibody.

    Techniques: Mouse Assay, Mutagenesis, Staining, Quantitation Assay, TUNEL Assay, Two Tailed Test

    Sec61a1 is a conserved β-cell gene whose mutation is associated with diabetes due to hypoinsulinemia. A : Amino acid comparison of mouse Sec61α1 residues 330–360 to the paralog mSec61α2 and orthologs (h, human; x, Xenopus ; y, yeast; b, bacteria). The asterisk shows Tyr344. B : Fasted plasma glucose and insulin were analyzed in Sec61a1 +/+ (wild type), Sec61a1 +/Y344H (HET), and Sec61a1 Y344H/Y344H (MUT) mice, aged 12 weeks, fed HFD. Males ( n = 4–10); females ( n = 9–13). Values shown are means ± SEM. Statistical assessments were made using a two-tailed Student t test assuming unequal variances (versus sex-matched wild-type littermates); * P

    Journal: Diabetes

    Article Title: A Point Mutation in Sec61?1 Leads to Diabetes and Hepatosteatosis in Mice

    doi: 10.2337/db08-1362

    Figure Lengend Snippet: Sec61a1 is a conserved β-cell gene whose mutation is associated with diabetes due to hypoinsulinemia. A : Amino acid comparison of mouse Sec61α1 residues 330–360 to the paralog mSec61α2 and orthologs (h, human; x, Xenopus ; y, yeast; b, bacteria). The asterisk shows Tyr344. B : Fasted plasma glucose and insulin were analyzed in Sec61a1 +/+ (wild type), Sec61a1 +/Y344H (HET), and Sec61a1 Y344H/Y344H (MUT) mice, aged 12 weeks, fed HFD. Males ( n = 4–10); females ( n = 9–13). Values shown are means ± SEM. Statistical assessments were made using a two-tailed Student t test assuming unequal variances (versus sex-matched wild-type littermates); * P

    Article Snippet: Antibody combinations were as follows: rabbit anti-insulin (sc-9168; Santa Cruz Biotechnology) and AlexaFluor 594–conjugated chicken anti-rabbit antibody; goat anti-glucagon antibody (sc-97780; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated donkey anti-goat antibody; goat anti-SEC61A1 antibody (ab1327; Abcam) and AlexaFluor 488–conjugated donkey anti-goat antibody; anti-immunoglobulin heavy chain–binding protein (BIP) antibody (sc-1050; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated donkey anti-goat antibody; anti-C/EBP homologous protein (CHOP) antibody (sc-7351; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated goat anti-mouse antibody.

    Techniques: Mutagenesis, Mouse Assay, Two Tailed Test

    Transgenic rescue of diabetes in mutant mice by β-cell–specific expression of wild-type Sec61a1. A : Expression as assessed by qPCR of Sec61a1 in islets or liver of wild-type animals or animals transgenic for Sec61a1 driven by the rat insulin promoter (+/+, RIP-Sec61); n = 3 mice of each genotype, each measured in triplicate. Values shown are means ± SEM among animals of the same genotype. Statistical assessments were made using a single-factor (genotype) ANOVA; ** P

    Journal: Diabetes

    Article Title: A Point Mutation in Sec61?1 Leads to Diabetes and Hepatosteatosis in Mice

    doi: 10.2337/db08-1362

    Figure Lengend Snippet: Transgenic rescue of diabetes in mutant mice by β-cell–specific expression of wild-type Sec61a1. A : Expression as assessed by qPCR of Sec61a1 in islets or liver of wild-type animals or animals transgenic for Sec61a1 driven by the rat insulin promoter (+/+, RIP-Sec61); n = 3 mice of each genotype, each measured in triplicate. Values shown are means ± SEM among animals of the same genotype. Statistical assessments were made using a single-factor (genotype) ANOVA; ** P

    Article Snippet: Antibody combinations were as follows: rabbit anti-insulin (sc-9168; Santa Cruz Biotechnology) and AlexaFluor 594–conjugated chicken anti-rabbit antibody; goat anti-glucagon antibody (sc-97780; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated donkey anti-goat antibody; goat anti-SEC61A1 antibody (ab1327; Abcam) and AlexaFluor 488–conjugated donkey anti-goat antibody; anti-immunoglobulin heavy chain–binding protein (BIP) antibody (sc-1050; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated donkey anti-goat antibody; anti-C/EBP homologous protein (CHOP) antibody (sc-7351; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated goat anti-mouse antibody.

    Techniques: Transgenic Assay, Mutagenesis, Mouse Assay, Expressing, Real-time Polymerase Chain Reaction

    Failure to rescue hepatic steatosis in mutant mice by β-cell–specific expression of wild-type Sec61a1. A : Hematoxylin-eosin (H E, left), oil red O (middle), and Masson (right) staining of liver from Sec61a1 +/+ (+/+), Sec61a1 Y344H/Y344H (Y344H/Y344H), and Sec61a1 Y344H/Y344H , Tg[RIP-Sec61] (Y344H/Y344H, RIP-Sec61) mice. B : Hepatomegaly was assessed by dissecting and weighing liver from four males each of Sec61a1 +/+ (+/+), Sec61a1 Y344H/Y344H (Y344H/Y344H), and Sec61a1 Y344H/Y344H , Tg[RIP-Sec61] (Y344H/Y344H, RIP-Sec61) mice. The liver weight is expressed as a percentage of total body weight (mean ± SEM). Statistical assessments were made using a two-tailed Student t test assuming unequal variances (versus wild-type littermates); ** P

    Journal: Diabetes

    Article Title: A Point Mutation in Sec61?1 Leads to Diabetes and Hepatosteatosis in Mice

    doi: 10.2337/db08-1362

    Figure Lengend Snippet: Failure to rescue hepatic steatosis in mutant mice by β-cell–specific expression of wild-type Sec61a1. A : Hematoxylin-eosin (H E, left), oil red O (middle), and Masson (right) staining of liver from Sec61a1 +/+ (+/+), Sec61a1 Y344H/Y344H (Y344H/Y344H), and Sec61a1 Y344H/Y344H , Tg[RIP-Sec61] (Y344H/Y344H, RIP-Sec61) mice. B : Hepatomegaly was assessed by dissecting and weighing liver from four males each of Sec61a1 +/+ (+/+), Sec61a1 Y344H/Y344H (Y344H/Y344H), and Sec61a1 Y344H/Y344H , Tg[RIP-Sec61] (Y344H/Y344H, RIP-Sec61) mice. The liver weight is expressed as a percentage of total body weight (mean ± SEM). Statistical assessments were made using a two-tailed Student t test assuming unequal variances (versus wild-type littermates); ** P

    Article Snippet: Antibody combinations were as follows: rabbit anti-insulin (sc-9168; Santa Cruz Biotechnology) and AlexaFluor 594–conjugated chicken anti-rabbit antibody; goat anti-glucagon antibody (sc-97780; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated donkey anti-goat antibody; goat anti-SEC61A1 antibody (ab1327; Abcam) and AlexaFluor 488–conjugated donkey anti-goat antibody; anti-immunoglobulin heavy chain–binding protein (BIP) antibody (sc-1050; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated donkey anti-goat antibody; anti-C/EBP homologous protein (CHOP) antibody (sc-7351; Santa Cruz Biotechnology) and AlexaFluor 488–conjugated goat anti-mouse antibody.

    Techniques: Mutagenesis, Mouse Assay, Expressing, Staining, Two Tailed Test

    Recombinant ACKR3 and AVPR1A form heteromeric complexes. ( a ) Typical PLA images for the detection of individual receptors and receptor–receptor interactions in HEK293T cells transfected with DNA encoding HA- or FLAG-tagged receptors. Ctrl: cells transfected with pcDNA. Images show merged PLA/DAPI signals acquired from z -stack images ( n = 10; thickness 1 µm, bottom to top). Scale bars, 10 µm. ( b ) Quantification of PLA signals per cell as in ( a ). n = 3 independent experiments with n = 10 images per condition and experiment. ( c ) HEK293T cells expressing HA-AVPR1A and FLAG-ACKR3 were lysed (input), the lysate was immunoprecipitated (IP) with anti-HA, followed by immunoblotting (IB) to detect HA-AVPR1A (left) and FLAG-ACKR3 (right) in the IP samples. IP control: precipitate after incubation of cell lysates with IgG-coupled resin. ( d , e ) Intermolecular BRET assays. Cells were co-transfected with AVPR1A-hRluc plus EYFP (open circles) or ACKR3-EYFP (grey squares) at various acceptor : donor ratios ( d ) and with increasing amounts of AVPR1A-hRluc and ACKR3-EYFP at a constant ratio of 1 : 10 ( e ).

    Journal: Open Biology

    Article Title: Identification and functional characterization of arginine vasopressin receptor 1A : atypical chemokine receptor 3 heteromers in vascular smooth muscle

    doi: 10.1098/rsob.170207

    Figure Lengend Snippet: Recombinant ACKR3 and AVPR1A form heteromeric complexes. ( a ) Typical PLA images for the detection of individual receptors and receptor–receptor interactions in HEK293T cells transfected with DNA encoding HA- or FLAG-tagged receptors. Ctrl: cells transfected with pcDNA. Images show merged PLA/DAPI signals acquired from z -stack images ( n = 10; thickness 1 µm, bottom to top). Scale bars, 10 µm. ( b ) Quantification of PLA signals per cell as in ( a ). n = 3 independent experiments with n = 10 images per condition and experiment. ( c ) HEK293T cells expressing HA-AVPR1A and FLAG-ACKR3 were lysed (input), the lysate was immunoprecipitated (IP) with anti-HA, followed by immunoblotting (IB) to detect HA-AVPR1A (left) and FLAG-ACKR3 (right) in the IP samples. IP control: precipitate after incubation of cell lysates with IgG-coupled resin. ( d , e ) Intermolecular BRET assays. Cells were co-transfected with AVPR1A-hRluc plus EYFP (open circles) or ACKR3-EYFP (grey squares) at various acceptor : donor ratios ( d ) and with increasing amounts of AVPR1A-hRluc and ACKR3-EYFP at a constant ratio of 1 : 10 ( e ).

    Article Snippet: A total of 50 µg of rabbit anti-AVPR1A (Bioss BS-11598R), mouse anti-HA (Bioss bsm-50131M) or anti- rabbit IgG (AbCam Ab27478) were incubated with 50 µl of Amino Link Plus coupling resin for 180 min at room temperature.

    Techniques: Recombinant, Proximity Ligation Assay, Transfection, Expressing, Immunoprecipitation, Incubation, Bioluminescence Resonance Energy Transfer

    ACKR3 and AVPR1A form heteromeric complexes in hVSMCs. ( a ) Representative PLA images for the detection of individual receptors, receptor–receptor complexes and pMLC2 in hVSMC. PLA for pMLC2 was performed in non-permeabilized cells (pMLC2, bottom left) and permeabilized cells (pMLC2, permeabilized cells, bottom right). All other PLAs were performed in non-permeabilized cells. Ctrl: omission of one primary antibody. Images show merged PLA/DAPI signals acquired from z -stack images ( n = 10; thickness 1 µm, bottom to top). Scale bars, 10 µm. ( b ) Quantification of PLA signals per cell as in ( a ). n = 4 independent experiments with n = 10 images per condition and experiment. ( c ) Three-dimensional representations of ACKR3 : AVPR1A interactions in hVSMC. Deconvolved images were generated from z -stack images ( n = 20; thickness: 0.5 µm, bottom to top). Images show merged PLA/DAPI signals. ( d–g ) hVSMCs were lysed (=input) and AVPR1A was immunoprecipitated (IP) followed by immunoblotting (IB) to detect AVPR1A ( d ), CXCR4 ( e ), ACKR3 ( f ) and β 2 -AR ( g ) in the IP samples. IP control: precipitate after incubation of cell lysates with IgG-coupled resin. Images are representative of n = 4 independent experiments.

    Journal: Open Biology

    Article Title: Identification and functional characterization of arginine vasopressin receptor 1A : atypical chemokine receptor 3 heteromers in vascular smooth muscle

    doi: 10.1098/rsob.170207

    Figure Lengend Snippet: ACKR3 and AVPR1A form heteromeric complexes in hVSMCs. ( a ) Representative PLA images for the detection of individual receptors, receptor–receptor complexes and pMLC2 in hVSMC. PLA for pMLC2 was performed in non-permeabilized cells (pMLC2, bottom left) and permeabilized cells (pMLC2, permeabilized cells, bottom right). All other PLAs were performed in non-permeabilized cells. Ctrl: omission of one primary antibody. Images show merged PLA/DAPI signals acquired from z -stack images ( n = 10; thickness 1 µm, bottom to top). Scale bars, 10 µm. ( b ) Quantification of PLA signals per cell as in ( a ). n = 4 independent experiments with n = 10 images per condition and experiment. ( c ) Three-dimensional representations of ACKR3 : AVPR1A interactions in hVSMC. Deconvolved images were generated from z -stack images ( n = 20; thickness: 0.5 µm, bottom to top). Images show merged PLA/DAPI signals. ( d–g ) hVSMCs were lysed (=input) and AVPR1A was immunoprecipitated (IP) followed by immunoblotting (IB) to detect AVPR1A ( d ), CXCR4 ( e ), ACKR3 ( f ) and β 2 -AR ( g ) in the IP samples. IP control: precipitate after incubation of cell lysates with IgG-coupled resin. Images are representative of n = 4 independent experiments.

    Article Snippet: A total of 50 µg of rabbit anti-AVPR1A (Bioss BS-11598R), mouse anti-HA (Bioss bsm-50131M) or anti- rabbit IgG (AbCam Ab27478) were incubated with 50 µl of Amino Link Plus coupling resin for 180 min at room temperature.

    Techniques: Proximity Ligation Assay, Generated, Immunoprecipitation, Incubation

    FKN and CX3CR1 mediate WEHI 274.1 recruitment to injured atherosclerotic carotid arteries. ( A ) WEHI 274.1 adhesion to mechanically injured atherosclerotic carotids was studied over 30 minutes in the presence of an IgG control antibody (black bars) or a function blocking anti-FKN antibody (white bars). ( B ) Representative intravital microscopic images from mouse carotid arteries at baseline (left) and 15 minutes (right) after injury of the atherosclerotic vascular wall, pretreated with either an anti-FKN antibody (upper row) or an isotype IgG control antibody (lower row). WEHI 274.1 cells were stained with DCF (green). Bars, 50 µm. n = 4–8, *p

    Journal: PLoS ONE

    Article Title: Fractalkine Is Expressed in Early and Advanced Atherosclerotic Lesions and Supports Monocyte Recruitment via CX3CR1

    doi: 10.1371/journal.pone.0043572

    Figure Lengend Snippet: FKN and CX3CR1 mediate WEHI 274.1 recruitment to injured atherosclerotic carotid arteries. ( A ) WEHI 274.1 adhesion to mechanically injured atherosclerotic carotids was studied over 30 minutes in the presence of an IgG control antibody (black bars) or a function blocking anti-FKN antibody (white bars). ( B ) Representative intravital microscopic images from mouse carotid arteries at baseline (left) and 15 minutes (right) after injury of the atherosclerotic vascular wall, pretreated with either an anti-FKN antibody (upper row) or an isotype IgG control antibody (lower row). WEHI 274.1 cells were stained with DCF (green). Bars, 50 µm. n = 4–8, *p

    Article Snippet: Rat IgG2b and rabbit IgG, used as negative controls, were from Abcam and Dako, respectively ( ).

    Techniques: Blocking Assay, Staining