A0564

Manufactured by Merck Group

A0564 is a laboratory equipment product from Merck Group. It is a precision instrument designed for scientific research and analysis applications. The core function of A0564 is to provide accurate measurements and data collection for various experiments and studies. Further details on the intended use or specific features of this product are not available.

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Lab products found in correlation

Lsm 700, by Zeiss (2 mentions) Sigma b5002, by Merck Group (1 mentions) Ov tl 12 30, by Agilent Technologies (1 mentions) Alexa fluor conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions) Mab1637, by Merck Group (1 mentions) Ab34771, by Abcam (1 mentions) Fsx100 all in one microscope, by Olympus (1 mentions) Axioobserver z1 inverted microscope, by Olympus (1 mentions) Ma5 15257, by Thermo Fisher Scientific (1 mentions)

3 protocols using a0564

Islet Morphometric Analysis and Metabolic Assays

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Insulin measurements from plasma and pancreatic extracts were measured by radioimmunoassay (RIA) (Millipore), and blood glucose and luciferase assays were measured as described previously (Poy et al., 2009 (link)). Islet morphometric analysis after intraperitoneal injections of BrdU on four consecutive days (50 μg/g BW, Sigma B5002) was performed on 8 μm sections of paraffin-embedded pancreas approximately 150–200 μm apart. Sections were dewaxed, washed, and stained for insulin (Dako A0564), glucagon (Millipore AB932), BrdU (Abcam ab6326), Ki-67 (Dako), or TUNEL (Roche cat. no.12156792910). Cell numbers from all islets in 3–7 sections were counted with ImageJ software from 20X images obtained using a Zeiss LSM700 (Schneider et al., 2012 (link)). β cell mass was measured as the ratio of insulin-positive cell area to the total tissue area, multiplied by the weight of the pancreas using Imaris software (Bitplane). In vivo insulin release and glucose (GTT) or insulin (ITT) tolerance tests were performed following a 6 hr fast and injected intraperitoneally with either glucose (2 g/kg BW) or insulin (0.75 U/kg BW).

Tattikota S.G., Rathjen T., McAnulty S.J., Wessels H.H., Akerman I., van de Bunt M., Hausser J., Esguerra J.L., Musahl A., Pandey A.K., You X., Chen W., Herrera P.L., Johnson P.R., O’Carroll D., Eliasson L., Zavolan M., Gloyn A.L., Ferrer J., Shalom-Feuerstein R., Aberdam D, & Poy M.N. (2014). Argonaute2 Mediates Compensatory Expansion of the Pancreatic β Cell. Cell Metabolism, 19(1), 122-134.

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Immunostaining of Mammalian Pancreas

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Samples – both whole mount rodent pancreas and 1 mm-thick human specimens - were incubated for 2 days at room temperature with primary antibody (1:100 dilution) in PBST, washed 3 times with PBST, and incubated for 2 days at room temperature with AlexaFluor-conjugated secondary antibody (Jackson, 1:100 dilution) in PBST. For immunostaining of samples derived from frozen clinical specimens, an identical procedure was applied except preceded by overnight blocking in 3% Normal Donkey Serum + 0.3% Triton-X in PBS at room temperature.
List of labeling reagents used in this study:

anti-Parvalbumin (rabbit, Abcam ab11427)

anti-Insulin (guinea pig, Dako A0564)

anti-Tuj1 (mouse, Millipore MAB1637)

anti-Col4 (rabbit, Millipore MAB8201)

anti-GFP 488 (rabbit, Thermo-Fisher A21311)

anti-Glucagon (guinea pig, Takara M182)

anti-CD31 (rabbit, Abcam ab28364)

anti-Laminin (rabbit, Millipore AB2034)

anti-Somatostatin (rabbit, Peninsula T-4103)

anti-melanA (mouse, Dako A103)

anti-CK7 (mouse, Dako OV-TL 12/30)

anti-CK20 (mouse, Dako Ks20.8)

Propidium Iodide (Cell Signaling Technologies 4087 S)

DyLight-488 Lectin (Vector DL-1174)

Hsueh B., Burns V.M., Pauerstein P., Holzem K., Ye L., Engberg K., Wang A.C., Gu X., Chakravarthy H., Arda H.E., Charville G., Vogel H., Efimov I.R., Kim S, & Deisseroth K. (2017). Pathways to clinical CLARITY: volumetric analysis of irregular, soft, and heterogeneous tissues in development and disease. Scientific Reports, 7, 5899.

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Immunohistochemical Analysis of Mouse Tissues

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Mouse tissues were collected and processed for paraffin sections as previously described [23] . Primary antibodies and dilution for immunostaining or immunofluorescence were: β-galactosidase (1:50, Abcam #ab9361), insulin (1:500, Dako #A0564), Cre (1:100, Millipore # 69050-3), RFP for tdTomato (rabbit polyclonal, 1:100, Abcam #ab34771), RFP for tdTomato (mouse monoclonal, 1:250, ThermoFisher #MA5-15257), glucagon (1:500, Invitrogen #18-0064), Nkx6.1 (1:50, Santa Cruz Biotechnology #sc-15027), and Amylase (1:800, Cell Signaling #3796). Images were acquired on a Nikon Coolscope digital microscope, a Zeiss Axio Observer Z1 inverted microscope, or an Olympus FSX100 all-in-one microscope.

Ye R., Wang M., Wang Q.A., Spurgin S.B., Wang Z.V., Sun K, & Scherer P.E. (2016). Autonomous interconversion between adult pancreatic α-cells and β-cells after differential metabolic challenges. Molecular Metabolism, 5(7), 437-448.

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