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Guinea pig anti insulin antibody

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The guinea pig anti-insulin antibody is a laboratory reagent used in research applications. It is a specific antibody produced in guinea pigs that recognizes and binds to insulin, a hormone that regulates blood sugar levels. This antibody can be used in various analytical techniques to detect and quantify insulin in biological samples.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Polysine slides, by Avantor (2 mentions) Anti guinea pig alexa 546, by Thermo Fisher Scientific (2 mentions) Lsm 710, by Zeiss (2 mentions) Triton x 100, by Merck Group (1 mentions) Ab15309, by Abcam (1 mentions) Anti cd4, by BD (1 mentions) Prolong gold antifade mountant, by Thermo Fisher Scientific (1 mentions) Hoechst, by Thermo Fisher Scientific (1 mentions) Accuri c6 system, by BD (1 mentions) Perm wash buffer, by BD (1 mentions) Fluoview fv1000d, by Olympus (1 mentions) Mouse anti β catenin antibody, by BD (1 mentions) Alexa fluor 488 labeled goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Fluorescein conjugated secondary antibody, by Merck Group (1 mentions) Dapi 4 6 diamidino 2 phenylindole, by Merck Group (1 mentions) Blocking one, by Nacalai Tesque (1 mentions) Scmos camera, by Oxford Instruments (1 mentions) Goat anti mouse alexa 488, by Thermo Fisher Scientific (1 mentions) Goat anti mouse cf594, by Biotium (1 mentions)

Close spelling variants of this product

Polyclonal guinea pig anti-insulin antibody Guinea pig anti-swine insulin antibody Guinea pig anti-insulin Pig anti-insulin antibody Anti-insulin antibody Insulin antibody Anti-insulin Insulin Anti-TM

18 protocols using guinea pig anti insulin antibody

1

Quantifying Pancreatic Islet Beta Cells

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Immunohistochemical staining of insulin was performed in order to characterize the integrity of beta cells in Langerhans islets. For this, tissue sections showing the maximum larger surface of Langerhans islets were selected for comparisons between groups. Sections were firstly deparaffined, hydrated through a decreasing gradual ethanol series, immersed in tampon citrate (pH 6) for 40 min, and then incubated with a polyclonal Guinea Pig anti-insulin antibody diluted in 50 mM Tris-HCl, pH 7,6 (DAKO). After washing, sections were incubated with phosphatase conjugated anti-Guinea Pig antibody and the procedure of revelation was assisted by TechMate 500 machine. Finally, sections were stained with haematoxylin/eosin.
Ten pancreatic islets per rat were examined under the microscope and the number of insulin-positive and insulin-negative cells was counted by numbering their nuclei. Data are expressed as the percent of insulin immunopositive cells/total number of cells in each islet.
Missaoui S., Ben Rhouma K., Yacoubi M.T., Sakly M, & Tebourbi O. (2014). Vanadyl Sulfate Treatment Stimulates Proliferation and Regeneration of Beta Cells in Pancreatic Islets. Journal of Diabetes Research, 2014, 540242.
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2

Immunofluorescence detection of enterovirus and insulin

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Mock-infected and CV-B4-infected INS-1 cells were seeded on sterile glass slides in 24-well plates. The cells were then fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100 (Sigma-Aldrich). The presence of intracellular enteroviral capsid protein VP1 and insulin in INS-1 cells was investigated using an immunofluorescence assay as previously described [29 (link),33 (link)]. Briefly, permeabilized cells were incubated overnight with a mouse anti-VP1 antibody, clone 5D8/1 (DAKO, Les Ulis, France) and a guinea pig anti-insulin antibody (DAKO, Via Real, CA, USA). The respective secondary antibodies: Alexa Fluor™ 488-conjugated rabbit anti-mouse antibody and Alexa Fluor™ 594-conjugated goat anti-guinea pig (Abcam, Cambridge, UK) were then applied sequentially for 1 h at room temperature. Cell nuclei were then stained with Hoescht dye solution, (InvitrogenTM) for 15 min. The slides were observed under a Leica DMi8 fluorescence microscope (Leica Microsystemes, Nanterre, France) at ×40, and positive cells were visualized with specific detector filters.
Nekoua M.P., Bertin A., Sane F., Gimeno J.P., Fournier I., Salzet M., Engelmann I., Alidjinou E.K, & Hober D. (2021). Persistence of Coxsackievirus B4 in Pancreatic β Cells Disturbs Insulin Maturation, Pattern of Cellular Proteins, and DNA Methylation. Microorganisms, 9(6), 1125.
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3

Pancreatic Cryosectioning and Immunofluorescence

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Pancreata were fixed in 4% paraformaldehyde for 72 hr, dehydrated in sucrose and mounted in OCT. Ten‐micron sections were cut on a Leica cryostat onto Polysine slides (VWR), air‐dried and fixed again for 10 min in acetone. Guinea‐pig‐anti‐insulin antibody (DAKO) was detected with anti‐guinea‐pig Alexa 546 (Molecular Probes, Eugene, OR). Anti‐CD4 (BD) was detected with anti‐rat Alexa 488 (Molecular Probes). GLUT‐1 in cells was detected using ab15309 (Abcam, Cambridge, UK) and Alexa 647 anti‐rabbit (Molecular Probes) after permeabilization with PBS‐Triton‐X. Nuclei were visualized with DAPI (Molecular Probes). The sections were viewed with a confocal microscope (Zeiss, Oberkochen, Germany) and processed using zen software.
Wallberg M., Recino A., Phillips J., Howie D., Vienne M., Paluch C., Azuma M., Wong F.S., Waldmann H, & Cooke A. (2017). Anti‐CD3 treatment up‐regulates programmed cell death protein‐1 expression on activated effector T cells and severely impairs their inflammatory capacity. Immunology, 151(2), 248-260.
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4

Multiplex Immunofluorescence of Pancreas

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Pancreas sections were stained for insulin, proinsulin, and glucagon after a standard triple indirect immunofluorescence staining. After deparaffinization and rehydration in descending ethanol concentrations, sections were exposed to heat-based antigen retrieval (citrate buffer). Staining was performed using a polyclonal guinea pig anti-insulin antibody (1:500; Dako, Carpinteria, CA), monoclonal mouse anti-proinsulin (DSHB clone GS-9A8, 1:50), and monoclonal mouse anti-glucagon (clone K79bB10, 1:300; Abcam) conjugated in-house to Alexa Fluor 647. Secondary antibodies included F(ab′)2 fragment of goat anti-guinea pig IgG conjugated to Alexa Fluor 488 (1:800; Jackson ImmunoResearch) and goat anti-mouse IgG (H+L) conjugated to Alexa Fluor 555 (1:1,000; Life Technologies, Grand Island, NY) incubated at room temperature for 30 min. Sections were counterstained with Hoechst (1:400; Life Technologies) for 10 min and then mounted with ProLong Gold Antifade Mountant (Life Technologies).
Rodriguez-Calvo T., Zapardiel-Gonzalo J., Amirian N., Castillo E., Lajevardi Y., Krogvold L., Dahl-Jørgensen K, & von Herrath M.G. (2017). Increase in Pancreatic Proinsulin and Preservation of β-Cell Mass in Autoantibody-Positive Donors Prior to Type 1 Diabetes Onset. Diabetes, 66(5), 1334-1345.
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5

Insulin Expression Quantification in hESCs

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Single-cell suspensions of differentiated human ES cell cultures were obtained by dissociating cells with 0.25% trypsin, fixing with 2% paraformaldehyde and permeabilizing with BD Perm/Wash buffer (Becton Dickinson). The cells were incubated with guinea pig anti-insulin antibody (1:1000, Dako) for 30 min at room temperature and then stained with Alexa Fluor 488-conjugated goat antibody directed against guinea pig (1:800) for 30 min at room temperature. Flow cytometry was performed using an Accuri C6 system (Becton Dickinson).
Guo D., Liu H., Ruzi A., Gao G., Nasir A., Liu Y., Yang F., Wu F., Xu G, & Li Y.X. (2017). Modeling Congenital Hyperinsulinism with ABCC8-Deficient Human Embryonic Stem Cells Generated by CRISPR/Cas9. Scientific Reports, 7, 3156.
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6

Immunostaining of MIN6c4 cells

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MIN6c4 cells were fixed with 4% paraformaldehyde for 10 min and then permeabilized in PBS containing 0.2% Triton X-100 for 10 min at room temperature. The cells were blocked using Blocking One (Nacalai Tesque, Kyoto, Japan) for 60 min at room temperature followed by incubation with a primary antibody at 4°C overnight and then with a fluorescein-conjugated secondary antibody (1:400) and DAPI (4’, 6-diamidino-2-phenylindole) (Sigma-Aldrich, Saint Louis, MO, USA) for 60 min at room temperature. The stained cells were examined by confocal laser scanning microscopy using a FLUOVIEW FV1000D (Olympus, Tokyo, Japan). The following primary antibodies were used in this analysis: purified rabbit anti-TMEM59L antibody (1:400), guinea pig anti-insulin antibody (1:500, DAKO, Glostrup, Denmark), mouse anti-GM130 antibody (1:500, BD Transduction Laboratories, San Jose, CA, USA), and mouse anti-β-catenin antibody (1:1,000, BD Transduction Laboratories). The following secondary antibodies were used in this analysis: Alexa Fluor 488-labeled goat anti-rabbit IgG, Alexa Fluor 594-labeled goat anti-guinea pig IgG, and Alexa Fluor 647-labeled goat anti-mouse IgG1 (Life Technologies, Grand Island, NY, USA).
Kobayashi M., Yamato E., Tanabe K., Tashiro F., Miyazaki S, & Miyazaki J.I. (2016). Functional Analysis of Novel Candidate Regulators of Insulin Secretion in the MIN6 Mouse Pancreatic β Cell Line. PLoS ONE, 11(3), e0151927.
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7

Immunohistochemical Analysis of Insulin, PECAM, and NG2

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Tissues were fixed on Day 3 with 100% Methanol. We then washed and stained the tissues for insulin, CD31(PECAM), and neuron glial antigen 2. All antibodies were diluted in PBS + 0.1% saponin + 2% BSA, and tissues were washed three times with PBS + 0.1% saponin for 10 mins between each antibody incubation. Insulin/PECAM/NG2 panels used 1:1000 dilution of guinea pig anti insulin antibody (Dako, Santa Clara, CA, USA), 1:50 dilution of biotinylated mouse monoclonal anti CD31 primary antibody (BD Biosciences, San Jose, CA, USA ), and 1:100 dilution of rabbit anti-neuron-glial antigen 2 (NG2; MilliporeSigma, Burlington, MA, USA). Secondary antibodies were used at a concentration 1:200 for goat anti-rabbit (Jackson ImmunoResearch, West Grove, PA) and goat anti-guinea pig (Abcam, Cambridge, UK) antibodies and 1:500 dilution of streptavidin antibody conjugated to CY2 (Jackson ImmunoResearch, West Grove, PA). For Figures 1, 3, and 4, images were acquired using 4x and 10x objectives on an inverted microscope (Nikon eclipse Ti2) coupled with a Photometrics CoolSNAP EZ camera or an Andor Zyla sCMOS camera. Confocal images in Figure 5 were acquired using the 10x and 20x objectives on a Ziess LSM 710 coupled with an Axio Observer microscope.
Dolan R., Lampejo A., Santini-González J., Hodges N.A., Phelps E.A, & Murfee W.L. (2022). A Novel Ex Vivo Method for Investigating Vascularization of Transplanted Islets. Journal of vascular research, 59(4), 229-238.
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8

Fluorescence Imaging of MIP-Timer Mice

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Transgenic MIP-Timer embryos were killed, and macroscopic appearance and fluorescence of the MIP-Timer mice were examined with a fluorescent dissecting microscope. For histological analysis, tissues from 3-week-old MIP-Timer mice were fixed in 4% paraformaldehyde in PBS at 4°C, washed in PBS alone, then immersed into sucrose solution in PBS overnight at 4°C. The next day, the tissues were embedded and frozen in Tissue-Tek O.C.T. Compound (Sakura). Tissues were sectioned at 6-μm thickness, permeabilized with 0.1% Triton X-100, and incubated with guinea pig anti-insulin antibody (Dako, Carpinteria, CA) diluted 1:2,000 in PBS and then visualized by using Alexa Fluor 647 anti-guinea pig IgG (Molecular Probes, Eugene, OR).
Miyatsuka T., Matsuoka T.A., Sasaki S., Kubo F., Shimomura I., Watada H., German M.S, & Hara M. (2014). Chronological Analysis With Fluorescent Timer Reveals Unique Features of Newly Generated β-Cells. Diabetes, 63(10), 3388-3393.
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9

Immunofluorescent Characterization of 3D Liver and Pancreatic Tissues

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Liver spheroids and pancreatic islet microtissues were embedded in Tissue-Tek® O.C.T. compound (Sakura Finetek, Torrance, CA, USA) for immunohistochemistry. Central cryostat sections of 12 µm were fixed in acetone at −20 °C for 10 min, washed with PBS and blocked with 10% (v/v) goat serum in PBS for 20 min. The pancreatic islet microtissues were immunostained with guinea pig anti-insulin antibody (Dako, Glostrup, Denmark) and mouse anti-glucagon (Abcam, Cambridge, UK) overnight, washed with PBS and, subsequently, developed by goat anti-guinea pig CF594 (Biotium, Fremont, CA, USA) and goat anti-mouse Alexa-488 (Life Technologies, Carlsbad, Ca, USA) for 45 min. DAPI was added for nuclei staining. The same procedure was carried out for the liver spheroids using mouse anti-cytokeratin 8/18 (Santa Cruz, Heidelberg, Germany), rabbit anti-vimentin (Santa Cruz), goat anti-albumin FITC (Bethyl Laboratories, Montgomery, Tx, USA) and mouse anti-Cyp3A4 (Santa Cruz) as primary antibodies and goat anti-mouse Alexa-488 (Life Technologies, Carlsbad, Ca, USA), goat anti-rabbit CF594 (Biotium) or goat anti-mouse CF594 (Biotium) as secondary antibodies. Images were obtained using a Keyence fluorescence microscope
Bauer S., Wennberg Huldt C., Kanebratt K.P., Durieux I., Gunne D., Andersson S., Ewart L., Haynes W.G., Maschmeyer I., Winter A., Ämmälä C., Marx U, & Andersson T.B. (2017). Functional coupling of human pancreatic islets and liver spheroids on-a-chip: Towards a novel human ex vivo type 2 diabetes model. Scientific Reports, 7, 14620.
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10

Pancreatic Beta Cell Analysis in DJ-1 KO Mice

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After four weeks of MLDS treatment, pancreata of wild type and DJ-1 KO mice were isolated and fixed in 4% paraformaldehyde for 24 h. Evenly spaced 10 μm sections were used to determine the beta cell area by staining them for insulin using the polyclonal guinea pig anti-Insulin antibody (DAKO). As secondary antibody we used goat anti-guinea pig conjugated with Alexa-Fluor-555 (Molecular Probes). DAPI (Sigma) was used to stain cell nuclei. Relative insulin-positive area was determined by quantification of the cross-sectional insulin-positive area divided by the cross-sectional area of the whole pancreatic section (nuclei area) and presented as percentage of control. Co-staining of DJ-1 and insulin in pancreatic sections was performed using rabbit anti-DJ-1 (1:100, [23 (link)]) and guinea pig anti-Insulin (1:300 dilution, DAKO).
Jain D., Weber G., Eberhard D., Mehana A.E., Eglinger J., Welters A., Bartosinska B., Jeruschke K., Weiss J., Päth G., Ariga H., Seufert J, & Lammert E. (2015). DJ-1 Protects Pancreatic Beta Cells from Cytokine- and Streptozotocin-Mediated Cell Death. PLoS ONE, 10(9), e0138535.
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