Late trophozoite-stage (24–36 hr post invasion [hpi]) P. falciparum-infected erythrocytes were lysed in 0.09% saponin in 5 mM Tris pH 7.5 and washed three times in PBS to remove haemoglobin. Following centrifugation, the parasite pellet was solubilized by sonication in 0.25% (v/v) Triton X-100 or 1% (v/v) ASB-14 (3-(tetradecanoylamidopropyl dimethylammonio) propane 1-sulfonate), the latter because it is often used for solubisation of proteins for 2D electrophoresis), then incubated with mixing at 4°C for 30 min. Insoluble material was pelleted (14 000 g for 30 min at 4°C). The supernatants were electrophoresed on NativePAGE Novex 3–12% Bis-Tris protein gels as per manufacturer’s instructions (Invitrogen) and transferred to PVDF for Western blotting. Bound antibody probes were detected with LiCor Odyssey Fc infrared imager followed by analysis with ODYSSEY v1.2 software.
Licor odyssey fc infrared imager
Manufactured by LI COR
The LiCor Odyssey Fc infrared imager is a device designed for detection and quantification of fluorescent and chemiluminescent signals in a variety of applications, including Western blotting, gel imaging, and in-cell and in-tissue fluorescence analysis.
Automatically generated - may contain errors
Lab products found in correlation
Nativepage novex 3 12 bis tris protein gels, by Thermo Fisher Scientific (1 mentions)
Mouse anti flag m2 antibody, by Merck Group (1 mentions)
Pierce g2 fast blotter, by Thermo Fisher Scientific (1 mentions)
Near infrared secondary antibody, by LI COR (1 mentions)
Goat anti myoc antibody, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti gluc antibody, by New England Biolabs (1 mentions)
Odyssey blocking buffer, by LI COR (1 mentions)
Mouse anti β actin, by Merck Group (1 mentions)
2 protocols using licor odyssey fc infrared imager
1
Solubilization and Native Fractionation of Plasmodium falciparum Proteins
2
Quantitative Western Blot Analysis of Secreted MYOC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For all Western blot-related experiments on secreted MYOC, 2% FBS containing media was used to minimize gel warping and epitope masking due to excessive FBS/BSA. Conditioned media (20 μL) was denatured in either reducing or nonreducing Laemmli buffer at 95°C for 5 minutes and loaded onto a Tris-Gly SDS-PAGE gel. Proteins were separated and transferred onto a nitrocellulose membrane using a Pierce G2 Fast Blotter (Thermo Scientific). Membranes then were blocked in Odyssey blocking buffer (LI-COR, Lincoln, NE, USA), and probed with either a goat anti-MYOC antibody (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), a rabbit anti-GLuc antibody (1:2000; NEB), a mouse anti-FLAG M2 antibody (1:2000, Sigma-Aldrich Corp.), a rabbit anti-GRP94 (1:1500, GeneTex, Irvine, CA, USA), or a mouse anti-β-actin (1:10,000, Sigma-Aldrich Corp.) primary antibody followed by an appropriate near-infrared secondary antibody (1:10,000; LI-COR). A LI-COR Odyssey Fc infrared imager (LI-COR) was used to image the blots, and the Image Quant software was used to quantify protein bands.
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!