Western blots were performed as previously described [10 (link)]. Antibodies used were specific to IMP1 (Santa Cruz), IMP2/p62 [6 (link), 10 (link)] detecting both isoforms, IMP3 (Proteintech), and α-tubulin (#T9026, Sigma, Germany). IRDye680-conjugated anti-rabbit IgG (LiCor Bioscience, Germany) and IRDye800-conjugated anti-mouse IgG (LiCor) were used as secondary antibodies. Signal intensities were determined by using the Odyssey infrared imaging system (LiCor).
Irdye 800 conjugated anti mouse igg
Manufactured by LI COR
Sourced in Germany
The IRDye 800-conjugated anti-mouse IgG is a secondary antibody that binds to mouse immunoglobulin G (IgG) and is labeled with the IRDye 800 fluorescent dye. It can be used for detection and quantification of mouse IgG in various immunoassay applications.
Automatically generated - may contain errors
Lab products found in correlation
Irdye680 conjugated anti rabbit igg, by LI COR (4 mentions)
Odyssey blocking buffer, by LI COR (4 mentions)
Odyssey infrared imaging system, by LI COR (2 mentions)
Odyssey infrared scanner, by LI COR (2 mentions)
Alexa 680 conjugated anti rabbit igg, by Thermo Fisher Scientific (2 mentions)
Image studio software, by LI COR (2 mentions)
α tubulin, by Merck Group (1 mentions)
Mouse anti c myc, by Thermo Fisher Scientific (1 mentions)
Odyssey clx infrared imager, by LI COR (1 mentions)
Ripa buffer, by Thermo Fisher Scientific (1 mentions)
Bca assay, by Thermo Fisher Scientific (1 mentions)
Protease phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)
Nitrocellulose, by Bio-Rad (1 mentions)
Mouse anti flag antibody, by Merck Group (1 mentions)
Odyssey clx infrared imaging system, by LI COR (1 mentions)
Gradient gel, by Bio-Rad (1 mentions)
Irdye 680 conjugated anti mouse igg, by LI COR (1 mentions)
Irdye800 conjugated anti rabbit igg, by LI COR (1 mentions)
Atr n 19, by Santa Cruz Biotechnology (1 mentions)
P chk1 ser296, by Cell Signaling Technology (1 mentions)
7 protocols using irdye 800 conjugated anti mouse igg
1
Western Blot Analysis of IMP Proteins
2
Western Blot Protein Expression Analysis
3
Western Blot Analysis of Complemented Strains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Intracellular samples of the complemented strains were subjected to SDS-PAGE followed by Western blotting. Immunoblots were probed with mouse anti-c-Myc (Thermo; 1:500), rabbit anti-TgGRA7 (1:5,000–John Boothroyd Lab), in Odyssey LI-COR blocking buffer (LI-COR Biosciences), followed by incubation with IRDye 680-conjugated anti-rabbit IgG and IRDye 800-conjugated anti-mouse IgG (LI-COR), each at 1:20,000 in PBS containing 0.5% BSA. The blots were washed in PBS and scanned using an Odyssey CLx infrared imager (LI-COR). Images were processed using Image Studio software (LI-COR).
4
Immunoblotting of Extracellular Vesicles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunoblotting, EV samples were washed in PBS and immediately lysed in RIPA buffer (Thermo, Carlsbad, CA, USA) supplemented with phosphatase/protease inhibitor cocktail (Thermo). Protein concentrations quantitated by BCA assay (Thermo) were comparable in all the samples. Twenty micrograms of lysates were separated by SDS-PAGE and transferred on nitrocellulose (Bio-Rad, Hercules, CA, USA). The membrane was blocked with Odyssey blocking buffer (LI-COR Biosciences, Lincoln, NE, USA) for 1 hour before incubating with anti-ApoA1 (clone 5F4, Catalog #: 3350; Cell Signaling, Danvers, MA, USA), anti-ApoB11 (Catalog #: AF3620; R&D, Minneapolis, MN, USA), or anti-CD63 (Clone H5C6, Catalog #: 353013; Biolegend) antibodies overnight. Secondary Alexa Fluor 680-conjugated anti-Goat IgG (LI-COR Biosciences) and IRDye800-conjugated anti-Mouse IgG (LI-COR Biosciences) were incubated for 1 hour. Blots were imaged using an Odyssey Infrared Imaging System (LI-COR Biosciences).
5
Western Blot Analysis of Parasite Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein samples from 107 parasites were resolved by SDS–PAGE and transferred to polyvinylidene fluoride (PVDF) as previously described (Carey et al., 2004 (link)). Blots were probed with rabbit anti-TgMyoA antiserum at 1:1000, mouse anti-TgGRA8 at 1 μg/ml, and mouse anti-FLAG antibody (Sigma-Aldrich) at 1:7500 in PBS containing 0.5% (vol/vol) bovine serum albumin (BSA), followed by incubation with IRDye 680–conjugated anti-rabbit IgG, IRDye 800–conjugated anti-mouse IgG, and IRDye 680–conjugated goat anti-mouse IgG1 at (1:20,000; LI-COR Biosciences, Lincoln, NE), each at 1:20,000 in PBS containing 0.5% BSA. Blots were then washed in PBS and scanned using an Odyssey CLx Infrared Imaging System (LI-COR Biosciences). Images were processed using Image Studio software (LI-COR Biosciences).
6
Quantitative Protein Analysis by SDS-PAGE and Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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).
7
Western Blot Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were stimulated as indicated at 37°C, then lysed immediately in ice-cold buffer containing 1% Nonidet P-40, 10 μg/ml leupeptin and Pepstatin A each, 1 mM PMSF, and 1 mM sodium orthovanadate. Lysates were cleared of insoluble material by centrifugation at 16000 × g for 10 min at 4°C, then mixed with Laemmli buffer, boiled for 5 min and resolved on a 12% SDS-PAGE gel. Following overnight transfer to nitrocellulose membranes (10V, 4°C), blots were blocked for 1 hour with a 1:1 mix of PBS and Odyssey Blocking Buffer (LI-COR), incubated overnight at 4°C with mouse and rabbit primary antibodies, then incubated with Alexa 680-conjugated anti-rabbit IgG (Molecular Probes) and IRDye 800-conjugated anti-mouse IgG (LICOR) secondary antibodies. All antibody incubation steps were done in 1:1 PBS + Odyssey Blocking Buffer with 0.1% Tween-20. Membranes were imaged using an Odyssey infrared scanner (LI-COR), and densitometry done using ImageJ software (NIH).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!