Western of His-tagged protein.
I want to perform splenocyte restimulation culture for the estimation of Th1 and Th2 cytokines by ELISA. As each groups consists of 5 mice immunized with a test vaccine preparation; can splenocyte...
08 September 2019 4,493 2 View
My protein (64kDa) expressed in insoluble inclusion bodies. I tried to purify it by ion-exchange and affinity but unable to purify protein. So, want to try ammonium sulfate precipitation of...
02 March 2018 8,203 5 View
I want to amplify a gene whose end terminus sequence is in such a way that the after designing NotI restriction site inserting (reverse) primer; it forms primer dimer during PCR amplification. I...
08 September 2017 7,641 3 View
I am working on recombinant protein (Ig's) expressed in form of inclusion bodies. Upon giving certain treatments there is a up shift in protein band. If my protein is degrading during the process;...
10 November 2016 7,711 4 View
I have a protein with a 10xHis-MBP tag on the N-terminus. I'm fairly certain I am expressing my protein, but I'm not getting a signal on my western blot. Is there a possibility that the 10xHis tag...
25 February 2021 8,404 3 View
I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...
24 February 2021 8,514 3 View
Increasingly, more articles are being retracted because of duplicate or otherwise dubious Western blots. Consequently, journals are asking for original Western blots and authentication of...
23 February 2021 8,609 3 View
Hi Everyone, I am a bigginer in western blotting and I am trying to use it for validation of my proteomics data. I have: two regulated proteins with molecular mass of 40 and 73 kDa one control...
23 February 2021 6,671 4 View
I need help using the equation C1V1=C2V2 figuring out how much antibody to add to use for a western blot. The stock was origianlly 3.45 ng/ul but diluted 1:10 so now its 0.345 ng/ul. I need to run...
22 February 2021 9,901 3 View
I run western blot on HCT-116 cell lines, I got actin band like in the attached picture. The band size is not same. Can anyone help me explain this situation and how to overcome? Thank you your...
22 February 2021 6,937 3 View
I ran a western blot using 12% gel and probing for ERK/pERK (cell signaling antibodies). The green is total ERK, the Red is supposed to be phospho-ERK. I see 2 bands below the total ERK that...
22 February 2021 1,777 3 View
Hello, I am performing a western blot of my SARS-CoV-2 Spike-pseudotyped lentiviral particles. I denature my virus by adding concentrated virus to 2X Laemmli buffer at 1:1, then boiling for...
17 February 2021 4,844 4 View
Hi, so basically... I am trying to mimic a clinically observed condition wherein a frame-shift mutation happens only at one allele of the patients, giving rise to a truncated variant which evades...
17 February 2021 1,130 6 View
i have issues in expressing a chicken protein in BL21 cells with his tag. I do not see induction even at 18 degrees. Any suggestions
15 February 2021 8,939 3 View
