Are able to detect a Hu-single-chain variable fragment by western Blot?

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Is there a cell penetrating peptide that I can order/synthesis that can bind non-covalently to a protein cargo, and allow it to enter a cell (Hela)?

HI, I am trying to understand which cell penetrating peptide I can order/syth that will bind non covalently to a protein cargo (eg. GFP !) and then efficiently deliver it into a cell? Thanks

09 October 2017 4,650 2 View

Will a 10xHis-tag prevent anti-MBP from binding to MBP?

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

What's the concentration recommend for recombinant protein use in western blotting?

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

What is the best way to manage Western blots?

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

How to manage western blot for proteomics data validation?

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

Diltuions for a Western blot?

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

Can anyone help me how to get the similar bands in Actin?

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 see 2 random bands in my ERK/pERK blot? Has anyone else seen this?

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

Will SARS-CoV-2 Spike be broken into S1 and S2 by denaturing conditions?

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

Unable to detect a truncated protein variant, but I can observe a truncated cDNA from Sanger?

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

What is the cause of the banding obtained after purifying the PCR product with ExoSAPIT?

Hello We are working on a PCR product with an approximate size between 5000 and 7000 bp, we want to use it as a positive internal control, since we have sequenced it and it corresponds to the...

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Debasish Kumar Dey

Hello Ewan,

As per the best of my knowledge, scFv is a fusion protein and it is highly convenient to express the antigen-binding domain as similarly as a single peptide. So, I guess you can use HRP monoclonal conjugate secondary antibody for the detection.

Gertrudis Rojas

I always do this with an anti-tag mAb, because most scFvs are fused to a tag for detection or purification (His, myc, FLAF tag or any other). But I know people producing thir own anti-scFv antisera. Perhaps some commercial anti-Fab sera are crossreactive to several variable regions and thus useful, but I haven't tried.