Immunofluorescence staining was performed as previously reported10 (link). The following primary antibodies were used: anti-insulin (dilution 1:100, #4590; Cell Signaling Technology), anti-C-peptide (dilution 1:100, #4593S; Cell Signaling Technology), anti-PDX-1 (dilution 1:200, #5679; Cell Signaling Technology), anti-NKX6.1 (dilution 1:400, #54551; Cell Signaling Technology), anti-ICA (dilution 1:500, ab207750; Abcam plc.), anti-ZnT8 (dilution 1:500, 16169–1-AP, Proteintech, Chicago, IL, USA), anti-GAD65 (dilution 1:50, ab239372, Abcam plc.) and anti-PD-L1 (dilution 1:25, 17952–1-AP, Proteintech). Anti-rabbit Alexa 488 (A-11008; Thermo Fisher Scientific Inc.) for anti-C-peptide, PDX-1, ICA, ZnT8, GAD65 and PD-L1 antibodies, and anti-mouse Alexa 594 (A-11005; Thermo Fisher Scientific Inc.) for anti-insulin antibody were used as the secondary antibodies. 4′, 6-diamidino-2-phenylindole (DAPI) (P-36931; Thermo Fisher Scientific Inc.) was applied for nucleus staining.
Ab239372
Manufactured by Abcam
Sourced in United States
Ab239372 is a laboratory equipment product. It is designed for use in scientific research and experiments. The core function of this product is to [DESCRIPTION_NOT_AVAILABLE].
Automatically generated - may contain errors
Lab products found in correlation
Ab207750, by Abcam (2 mentions)
Anti pdx1, by Cell Signaling Technology (2 mentions)
Anti insulin, by Cell Signaling Technology (2 mentions)
A11008, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
A11005, by Thermo Fisher Scientific (1 mentions)
Anti c peptide, by Cell Signaling Technology (1 mentions)
Ab183685, by Abcam (1 mentions)
Ab217344, by Abcam (1 mentions)
Ab5905, by Merck Group (1 mentions)
Ab2723, by Abcam (1 mentions)
Ab216130, by Abcam (1 mentions)
Ab125011, by Abcam (1 mentions)
Ab133615, by Abcam (1 mentions)
β actin, by Merck Group (1 mentions)
Ab275377, by Abcam (1 mentions)
Ab92726, by Abcam (1 mentions)
Horseradish peroxidase conjugated goat anti rabbit sc 2004, by Santa Cruz Biotechnology (1 mentions)
Ab9722, by Abcam (1 mentions)
Ag 20b 0014, by Adipogen (1 mentions)
4 protocols using ab239372
1
Immunofluorescence Staining of Pancreatic Markers
2
Comprehensive Immunohistochemical Profiling of Pancreatic Cells
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Immunohistochemical staining was performed as previously reported56 (link) with the following antibodies: anti-insulin (dilution 1:100, #4590; Cell Signaling Technology), anti-PDX-1 (dilution 1:50, #5679; Cell Signaling Technology), anti-PD-L1 (dilution 1:100, 17952–1-AP, Proteintech), anti-ICA (dilution 1:500, ab207750; Abcam plc.), anti-ZnT8 (dilution 1:500, 16169–1-AP, Proteintech), anti-GAD65 (dilution 1:2000, ab239372; Abcam plc.), anti-CD4 (dilution 1:1000, ab183685; Abcam plc.) and anti-CD8 (dilution 1:2000, ab217344; Abcam plc.).
3
Immunoblot analysis of NLRP3 and exosomal markers
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NLRP3 (AG-20B-0014, AdipoGen, CA, USA); IL1β (ab9722, Abcam, MA, USA); Anti-CD63 antibody (ab216130, Abcam, MA, USA); Anti-CD9 antibody (ab92726, Abcam, MA, USA), Anti-TSG101 antibody (ab125011, Abcam, MA, USA), Anti-Alix antibody (ab275377, Abcam, MA, USA), Anti-Calnexin antibody (ab133615, Abcam, MA, USA); Anti-PSD95 antibody (ab2723, Abcam, MA, USA); Anti-GAD65 antibody (ab239372, Abcam, MA, USA); Anti-Gephyrin antibody (ab181382, Abcam, MA, USA); Anti-vGLUT1 antibody (AB5905, Millipore, Burlington, MA, USA), GFP expressing plasmid (13031, Adgene, Watertown, MA, USA); β-actin (A5316, Sigma- Aldrich, MO, USA); horseradish peroxidase conjugated goat anti-rabbit (sc-2004, Santa Cruz Biotechnology, TX, USA) and horseradish peroxidase conjugated goat anti-mouse (sc-2005, Santa Cruz Biotechnology, TX, USA); human primary microglia (Cat # 1900; Sciencell research laboratory, CA, USA) and BV2 microglial cell line was received from Dr. Sanjay Maggirwar (University of Rochester Medical Center, Rochester, NY, USA).
4
Quantitative Protein Analysis of Thalamic GAD and GAT-1
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The thalamus (n = 6 per group) was homogenized and lysed using RIPA (50 mmol/L Tris•HCl pH8.0, 150 mmol/L NaCl, 1%SDS, 1 mmol/L PMSF). The protein concentration was quantified using the Bradford method. The protein was used for Western blotting with GAD65 (1:5,000, ab239372, Abcam, Cambridge, United Kingdom), GAD67 (1:1,000, ab213508, Abcam, Cambridge, UK), and GAT-1 (1:1,000, ab177483, Abcam, Cambridge, United Kingdom) primary antibody, incubated overnight at 4°C. After incubation, the membranes were washed thrice with Tris-buffered saline containing Tween 20 and then incubated with the secondary antibody (1:10,000) at room temperature for 1 h, and washed with TBST six times for 3 min each. The blots were visualized using enhanced chemiluminescence (WBKLS0500, Millipore). The electrochemiluminescence signals were detected using Total Lab Quant V11.5 (Newcastle upon Tyne, United Kingdom). β-Actin (A20120A0702, BioTNT) was used as an internal control to validate the amount of protein loaded onto the gels.
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