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Anti glucagon

Manufactured by Agilent Technologies
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Sourced in Denmark, Germany

Anti-glucagon is a laboratory equipment product designed to detect and measure the presence of glucagon, a hormone produced by the pancreas that plays a crucial role in regulating blood sugar levels. This product provides a reliable and accurate way to analyze glucagon levels in various biological samples.

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Lab products found in correlation

Anti insulin, by Agilent Technologies (5 mentions) Anti pdx1, by Abcam (2 mentions) Anti glut2, by Merck Group (2 mentions) Fluorescein isothiocyanate fitc or cy3 conjugated secondary antibodies, by Jackson ImmunoResearch (2 mentions) Anti p mst1, by Cell Signaling Technology (2 mentions) Anti mouse anti ki67, by BD (2 mentions) Anti bim, by Cell Signaling Technology (2 mentions) Nis elements software, by Nikon (2 mentions) Vectashield with 4 6 diamidino 2 phenylindole dapi, by Vector Laboratories (2 mentions) Mea53200, by Nikon (2 mentions) Foxo4, by Cell Signaling Technology (1 mentions) Foxo3, by Cell Signaling Technology (1 mentions) Anti foxo1, by Cell Signaling Technology (1 mentions) α tubulin, by Cell Signaling Technology (1 mentions) Foxo1, by Cell Signaling Technology (1 mentions) Anti phospho paxillin y118, by Thermo Fisher Scientific (1 mentions) Anti as160, by Cell Signaling Technology (1 mentions) Recombinant tnf α, by R&D Systems (1 mentions) Monoclonal anti actin, by Merck Group (1 mentions) Anti akt, by Cell Signaling Technology (1 mentions)

6 protocols using anti glucagon

1

Quantifying Pancreatic Foxo Expression

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Western blots used the primary antibodies directed against the following proteins: Foxo1, Foxo3, Foxo4, α-tubulin from Cell Signaling (#2880, 2497, 9472, and 2144). We fixed and processed tissues for immunohistochemistry as described (Kitamura et al., 2009 (link)). We applied perfused fixation, and antigen retrieval for nuclear transcription factor detection (Nacalai USA) (Talchai et al., 2012b (link)). We used anti-Foxo1 (Cell Signaling #2880), anti-insulin and anti-glucagon from Dako (#A056401-2, A056501-2) for primary antibodies and FITC-, Cy3-, and Alexa-conjugated donkey secondary antibodies (Jackson Immunoresearch Laboratories, and Molecular Probes)(Kitamura et al., 2009 (link)). Antibodies to Pcsk1, Pcsk2, Glut-2 and Pdx-1 were from EMD Millipore (#AB10553, AB15610, 07-1402, 061384). For β-cell morphometry, 4 pancreatic sections from 4 mice from each genotype were sampled 150 μm apart, and used for a two-tailed paired Student’s t-test analysis. Total pancreas area was captured at 100x, and immunostaining area at 200x (Talchai et al., 2012b (link)). The ratio of immunoreactive area/pancreas area was converted from pixel to mm2 and plotted as mm2.
Kim-Muller J.Y., Zhao S., Srivastava S., Mugabo Y., Noh H.L., Kim Y.R., Madiraju S.R., Ferrante A.W., Skolnik E.Y., Prentki M, & Accili D. (2014). Metabolic Inflexibility Impairs Insulin Secretion And Results In MODY-like Diabetes In Triple FoxO-deficient Mice. Cell metabolism, 20(4), 593-602.
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2

Recombinant Cytokine and Antibody Protocol

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Recombinant TNFα, CX3CL1 and anti-CX3CL1 were obtained from R&D Systems Europe Ltd (Abingdon, United Kingdom). Anti-phospho-paxillin (Y118) was obtained from Invitrogen (Carlsbad, CA); Anti-paxillin from Becton Dickinson (San Jose, CA); Anti-phospho-Akt (Ser473) and anti-phospho-ERK-1/2 (Thr202 and Tyr204) were purchased from New England Biolabs (Beverly, MA). Anti-ERK1/2, anti-Akt, anti-AS160 and anti-phospho-(Ser/Thr)-AS160 were obtained from Cell Signaling Technology (Beverly, MA). Anti-p65 subunit of NF-κB (C-20) was obtained from Santa Cruz Biotechnology (Santa Cruz, CA) and monoclonal anti-actin from Sigma. Anti-IRS-1 and IRS-2 were a gift from Dr. M.F. White (Children's Hospital, Harvard University, Boston, MA). Anti-insulin and anti-glucagon were from Dako (Glostrup, Denmark).
Rutti S., Arous C., Schvartz D., Timper K., Sanchez J.C., Dermitzakis E., Donath M.Y., Halban P.A, & Bouzakri K. (2014). Fractalkine (CX3CL1), a new factor protecting β-cells against TNFα. Molecular Metabolism, 3(7), 731-741.
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3

Immunohistochemical Analysis of Pancreatic Markers

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Tissue sections were heated in Tris-EDTA buffer to retrieve antigen epitopes, blocked by 10% normal goat serum and Avidin/Biotin blocking reagent (Vector Laboratories, Burlingame, CA) and stained with the following primary antibodies at 4°C overnight: anti-insulin (DAKO, Baar, Switzerland, #A0564), anti-CYR61 (Santa Cruz Biotechnology, Inc., Heidelberg, Germany, #sc13100), anti-glucagon (DAKO, #A0565), anti-CD31 (Neomarkers Inc, Fremont, CA, #RB-10333-P), anti-E-cadherin (BD Biosciences, Basel, Switzerland, #610182). Sections were incubated with biotinylated secondary antibodies followed by Vectastain ABC Kit (Vector Laboratories). DAB peroxidase substrate (Sigma-Aldrich, Buchs, Switzerland) was used to reveal the signal from antibody-peroxidase complex. Sections were counterstained with hematoxylin before mounting.
Huang Y.T., Lan Q., Ponsonnet L., Blanquet M., Christofori G., Zaric J, & Rüegg C. (2015). The matricellular protein CYR61 interferes with normal pancreatic islets architecture and promotes pancreatic neuroendocrine tumor progression. Oncotarget, 7(2), 1663-1674.
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4

Immunostaining of Chick/Quail Grafts

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Isolated grafts were fixed in 4% formaldehyde in PBS. Cryosections were cut at 5µm thickness. Immunohistochemical analysis of sectioned chick or quail tissue was as published (25 (link)). The following primary antibodies were applied overnight at 4°C: Anti-GFP (Invitrogen A11122, 1:200), Anti-smooth muscle actin (Sigma A2547 1:200), Anti-smooth muscle actin (Abcam Ab5694 1:200), Anti-sarcomeric myosin (MF20), QCPN (DSHB undiluted), 8F3 (DSHB 1:25), Anti-cytokeratin (Abcam Ab9377, 1:100), QH1 (DSHB, 1:200), Anti-insulin (Abcam Ab63820, 1:100) and Anti-glucagon (DAKO, A0565, 1:200). The following secondary antibodies were applied at a 1:500 dilution for 90 min at room temperature: Alexa fluor 488 or 568 Goat anti-rabbit (Invitrogen); Alexa fluor 488 or 568 Goat anti-mouse (Invitrogen). TOPRO-3 (Invitrogen T3605) at 1 µmol/L was applied with secondary antibody. Sections were imaged in Z-stacks using a LSM510 META Confocal with 0.4 µm optical slices. All IHC images presented in figures are Z-projections. The mesothelial layer was distinguished by morphology combined with cytokeratin staining. Nuclei within the mesothelial layer were manually identified and then subsequently identified as QCPN positive or negative.
Winters N.I., Williams A.M, & Bader D.M. (2014). Resident progenitors, not exogenous migratory cells, generate the majority of visceral mesothelium in organogenesis. Developmental biology, 391(2), 125-132.
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5

Immunofluorescence Analysis of Pancreatic Tissues

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Pancreatic tissues were processed as previously described 66 (link). In brief, mouse pancreases were dissected and fixed in 4% formaldehyde at 4°C for 12h before embedding in paraffin. Human and mouse 4-μm sections were deparaffinized, rehydrated and incubated overnight at 4°C with anti-insulin (Dako), anti-P-MST1 (Cell Signaling), anti-Bim (Cell Signaling), anti-PDX-1 (abcam), anti-glucagon (Dako), anti-glut2 (Chemicon) and anti-mouse anti-Ki67 (BD Pharmingen) antibodies followed by fluorescein isothiocyanate (FITC)- or Cy3-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA). Slides were mounted with Vectashield with 4′6-diamidino-2-phenylindole (DAPI) (Vector Labs). beta-cell apoptosis for mouse sections or primary islets cultured on ECM dishes was analyzed by the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) technique according to the manufacturer’s instructions (In Situ Cell Death Detection Kit, TMR red; Roche) and double stained for insulin. Fluorescence was analyzed using a Nikon MEA53200 (Nikon GmbH, Dusseldorf, Germany) microscope and images were acquired using NIS-Elements software (Nikon).
Ardestani A., Paroni F., Azizi Z., Kaur S., Khobragade V., Yuan T., Frogne T., Tao W., Oberholzer J., Pattou F., Conte J.K, & Maedler K. (2014). MST1 is a novel regulator of apoptosis in pancreatic beta-cells. Nature medicine, 20(4), 385-397.
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6

Immunofluorescence Analysis of Pancreatic Tissues

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Pancreatic tissues were processed as previously described 66 (link). In brief, mouse pancreases were dissected and fixed in 4% formaldehyde at 4°C for 12h before embedding in paraffin. Human and mouse 4-μm sections were deparaffinized, rehydrated and incubated overnight at 4°C with anti-insulin (Dako), anti-P-MST1 (Cell Signaling), anti-Bim (Cell Signaling), anti-PDX-1 (abcam), anti-glucagon (Dako), anti-glut2 (Chemicon) and anti-mouse anti-Ki67 (BD Pharmingen) antibodies followed by fluorescein isothiocyanate (FITC)- or Cy3-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA). Slides were mounted with Vectashield with 4′6-diamidino-2-phenylindole (DAPI) (Vector Labs). beta-cell apoptosis for mouse sections or primary islets cultured on ECM dishes was analyzed by the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) technique according to the manufacturer’s instructions (In Situ Cell Death Detection Kit, TMR red; Roche) and double stained for insulin. Fluorescence was analyzed using a Nikon MEA53200 (Nikon GmbH, Dusseldorf, Germany) microscope and images were acquired using NIS-Elements software (Nikon).
Ardestani A., Paroni F., Azizi Z., Kaur S., Khobragade V., Yuan T., Frogne T., Tao W., Oberholzer J., Pattou F., Conte J.K, & Maedler K. (2014). MST1 is a novel regulator of apoptosis in pancreatic beta-cells. Nature medicine, 20(4), 385-397.
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