Recommended volume per well for 1.0 mm mini gelġ0 μL for electrophoresis, 5 μL for blottingĥ μL for electrophoresis, 2. I have prepared my buffersagain but still its not. ~160, 120, 80, 60, 50, 40, 30, 20, 15, 10 kDaĮach protein standard contains a 6X His-tag enabling the detection with the InVision His-tag In-gel StainĬolorimetric or methods that detect phosphorylated proteins such as Pro-Q Diamond phosphoprotein gel stainsĬolorimetric or methods that detect glycosylated proteins such as Pro-Q Glycoprotein stain kits I am trying to transfer my proteins on PVDF membrane but only ladder transfers and i can see my protein on gel if stained after western blot. Peppermint Stick Phosphoprotein Molecular Weight StandardsĬand圜ane Glycoprotein Molecular Weight Standards Tagged- proteins in the 10 -250 kDa range contain a Strep-tag™ II sequence and can be detected on western blots using Strep-Tactin™ conjugates or an antibody against Strep-tag™ II sequenceĭirect visualization can be achieved through standard staining protocols (e.g., Coomassie, etc.) Tagged-proteins contain an integral Strep-tag™ II sequence and can be detected on western blots using Strep-Tactin™ conjugates or an antibody against Strep-tag™ II sequence PageRuler Unstained Broad Range Protein LadderĪccurate estimation across a broader range Recommended Volume per well for 1.0 mm gel (μL)Ĭolorimetric, NIR fluorescence (700nm channel, blue bands), RGB fluorescence (550 nm channel, orange bands) Now, some additional information: I run mini-gels all the time and never. Hi, I encountered something strange the other day with my western blot: after transfer, the pre-stained ladder (bio-rad) showed up fine, but there was no protein whatsoever on the blot stained with Ponceau. The last thing we have is that after a good transfer (25 mM Tris, 190 mM Gly, 0,1 SDS and 10 methanol for searching proteins of 120 KDa), with all bands of markers ok (prestained and kaleidoscope, both from BioRad) we have blocked with 5 milk (we have bought one with calcium and bifidus, don't know if there is a problem) in TBST. Ladder but no protein - (Aug/24/2009 ) Ladder but no protein. The strange thing here is that my ladder has actually completely transferred over and even passed through the membrane while my protein samples are all mostly stuck onto the gel. Visible monitoring of gel separation and transfer efficiency Hi everybody We are having problems with our western blots. In a nutshell Ive been having a great deal of problem transferring my proteins over to my nitrocellulose membrane for western blots. Spectra Multicolor Broad Range Protein Ladderīest multicolor prestained ladder for routine applicationsĤ colors for improved visualization during separation and transferĪnalysis of high molecular weight proteins
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |