We used antibodies for p-JNK, total JNK, p-ERK, total-ERK, AceH3K9, H3K917Me3, Bcl-XL, Caspase3, cleaved Caspase 3, Caspase 11, ILβ and cytochrome C (Cell Signaling Technology); β-actin and BAX (Santa Cruz Biotechnology, Inc.). Secondary antibodies were IRDye680 conjugated anti-mouse IgG and IRDye680 conjugated anti-goat IgG, IRDye800 conjugated anti-rabbit IgG and IRDye800 conjugated anti-goat IgG (Rockland, PA) (LICOR); TRITC- and FITC-conjugated donkey anti–mouse, anti–goat, and anti–rabbit (Jackson ImmunoResearch Laboratories, Inc.), and DAPI (Life-Tech); Antibodies CD68 (e-Bioscience), JC-1 staining kit (Thermo Fisher Scientific), and TUNEL assay kit (TMR Tunel staining kit, Roche) JNK-inhibitor SP600125 (Calbiochem) and ERK–inhibitor PD98059 (Calbiochem).
Irdye800 conjugated anti rabbit igg
Manufactured by LI COR
Sourced in United States
The IRDye800 conjugated anti-rabbit IgG is a secondary antibody conjugated with the IRDye800 fluorescent dye. It is designed for detecting and quantifying rabbit primary antibodies in various immunoassay applications.
Automatically generated - may contain errors
Lab products found in correlation
Irdye 680 conjugated anti mouse igg, by LI COR (4 mentions)
Odyssey infrared imaging system, by LI COR (2 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
P erk, by Cell Signaling Technology (1 mentions)
Total erk, by Cell Signaling Technology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Caspase 11, by Cell Signaling Technology (1 mentions)
Caspase 3, by Cell Signaling Technology (1 mentions)
P jnk, by Cell Signaling Technology (1 mentions)
Bcl xl, by Cell Signaling Technology (1 mentions)
Odyssey infrared image system, by LI COR (1 mentions)
Anti hdac1, by Abcam (1 mentions)
Anti hdac2, by Abcam (1 mentions)
Anti β actin, by Proteintech (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Anti hdac3, by Abcam (1 mentions)
Anti hdac8, by Abcam (1 mentions)
Anti cbx4, by Cell Signaling Technology (1 mentions)
Anti runx2, by Cell Signaling Technology (1 mentions)
Mouse monoclonal anti flag, by Merck Group (1 mentions)
6 protocols using irdye800 conjugated anti rabbit igg
1
Antibody-Based Protein Expression Analysis
2
Protein Expression Analysis in Surgical ACP
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Protein was extracted from the fresh ACP surgical specimens by lysing cells with protease inhibitor cocktail (Roche, USA). After measuring the protein content with a Bradford assay, 20 μg of protein was resolved by 10% SDS-PAGE and transferred to PVDF membranes. Membranes were blocked with 5% skim milk in TBST for 2 h at room temperature and incubated overnight with the following antibodies: anti-HAT, anti-HDAC1, anti-HDAC2, anti-HDAC3, anti-HDAC8 (1:1000, Abcam, USA), anti-CBX4, anti-Runx2, anti-histone (1:1000, Cell Signaling Technology, USA), and anti-β-actin (1:1000, Proteintech, USA). The specimens were then incubated with secondary antibodies, IRDye800-conjugated anti-rabbit IgG and IRDye680-conjugated anti-mouse IgG (1:15,000, LiCor, USA), for 1 h at room temperature to label the primary antibody. An Odyssey Infrared Image System (LiCor, USA) was used to analyze signal intensities. The densitometry results were first normalized to the density obtained for β-actin or histone and then compared with that of the control to obtain relative fold changes.
3
Standardized Western Blot Analysis
4
Western Blot Analysis of NLRP3 and NDV-NP
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DF1 cells were lysed with RIPA buffer supplemented with PMSF and phosphorylase inhibitors (Beyotime, Shanghai, China), and the protein concentration was tested with a Bradford assay (Bio-Rad, Hercules, CA, USA). Denatured protein (30 μg) was electrophoresed in a 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) gel and transferred to an NC membrane (GE, MA, USA). NC membranes were blocked and incubated with specific rabbit anti-NLRP3 (provided by Professor Zhangyong Ning of South China Agricultural University), anti-NDV-NP antibody, or rabbit anti-GAPDH antibody (Cat: ab181602, Abcam, UK) for 12 h at 4 °C. The primary antibody was removed, and the membrane was incubated with IRDye 800-conjugated anti-rabbit IgG (Cat: 926-32211, LI-COR Biosciences, Lincoln, NE, USA) for 45 min at 25 °C–30 °C. The membranes were visualized using an Odyssey infrared imaging system (LI-COR Biosciences, Lincoln, NE, USA).
5
Quantitative Protein Analysis by SDS-PAGE and Western Blot
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Total protein was extracted from exponentially growing cells at passage 8–10 and 40 g/mL were resolved by SDS-PAGE using a 4–15% gradient gel (Bio-Rad Laboratories). After transfer and blocking overnight at 4 °C in Odyssey Blocking Buffer (Li-Cor, Lincoln, NE, USA) proteins were detected by primary antibodies against BRCA1 [2A-9] (1:500, kindly provided by Stephen Smith, Leibnitz Institute, Jena, Germany), ATR [N-19] (Santa Cruz, St. Cruz, CA, USA, 1:1000), CHK1 [2G1D5] (Cell Signaling, Danvers, MA, USA, 1:750), RAD51 [14B4] (1:2.000, GeneTex, Irvine, CA, USA), MRE11A [12D7] (Abcam, Cambridge, UK, 1:500), pCHK1 [Ser296] (Cell Signaling, 1:1000), -actin [AC-74] (1:50.000, Sigma, St. Louis, MO, USA). Primary antibodies were detected with IRDYE 680 conjugated anti-mouse IgG, IRDYE 800 conjugated anti-rabbit IgG (Li-Cor, 1:7500), IRDYE 680 conjugated anti-rabbit IgG (Licor, 1:7.500 or 15.000) or IRDYE 800 conjugated anti-mouse IgG (Li-Cor 1:7.500 or 15.000). Quantitative and qualitative analysis was done by using Li-Cor Odyssey (Li-Cor, Lincoln, NE, USA).
6
Quantifying TRAIL Expression in AD-MSCs
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An in-cell Western method was performed as reported [25] . AD-MSC TRAIL and AD-MSC GFP were plated in triplicate for 24 hours. TRAIL presence was assayed by anti-TRAIL primary antibody (Peprotech, London, U.K.) followed by a secondary antibody conjugated to an infrared dye (IR dye 800 conjugated anti-rabbit IgG; LI-COR Biosciences, LI-COR Biosciences, Lincoln, NE). Secondary antibody and DRAQ5 Stain and Sap-phire700 normalizers (LI-COR Biosciences) were carried out simultaneously. Plates were scanned by Odyssey Infrared Imaging System (LI-COR Biosciences) in the 800-nm and 700nm channels. Signal was quantified using Odyssey Infrared Imaging System Application Software (LI-COR Biosciences).
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