Stijn Vdb

11 Questions 16 Answers 0 Followers

Questions related from Stijn Vdb

Concentrate iodixanol gradient ultracentrifugation fractions with 10kDa filter?

Hi, Can I concentrate iodixanol gradient ultracentrifugation fractions with a 10kDa filter? Iodixanol is about 1550 g/mol (= 1.55 kDa?), so it should work, but I've tried with 3kDa and did not...

16 July 2020 7,046 0 View

Appropriate lysis buffer for immunoprecipitation?

Hi, Which lysis buffer would you recommend to make a protein extract from tissue that does not interfere with immunoprecipitation afterwards? I'm currently using RIPA + protease inhibitors,...

21 October 2019 9,017 5 View

Slower migration on SDS-page in complex sample?

Hi all, Is it possible that the migration of my protein is delayed due to the complexity of the sample? When I run a western blot with 1) a protein extract from locust tissue and 2) hemolymph...

04 June 2019 10,098 10 View

BCA assay my sample is coloured?

Hi, I've performed a protein extraction on different insect tissues using a 2D DIGE lysis buffer (containing CHAPS, ureum, thioureum and DTT). When performing a BCA assay, my samples (which often...

02 April 2019 7,612 6 View

Secondary antibody gives same signal as primary?

Hi, After immunoprecipitation and elution, I check the eluate with western blot. If I incubate the blot ONLY with secondary antibody, it gives the same signal as incubatin with primary &...

16 January 2019 4,052 4 View

Western blot on immunoprecipitation eluate detects related protein?

Hello, So, first I perform immunoprecipitation. The eluate is then subjected to immunoblot analysis using an antibody against a related protein (equal size) and gives a band. How sure am I the...

14 November 2018 5,658 7 View

QPCR spike in normalization?

Can someone give me an easy example of how to normalize qPCR data against spike-in control? Thanks

06 March 2018 9,765 1 View

How to separate a complex fluid in a vesicular and non vesicular part

Hello, Is it possible to separate a complex fluid in a vesicular part (containing vesicles / exosomes) and a non - vesicular part, containing other proteins? Many thanks!

29 June 2017 5,684 3 View

What is the function of XBP buffer in exosome isolation?

Hello, What is the function of XBP buffer in exosome isolation? Does it solely bind to exosomes, or also other protein complexes?

08 May 2017 3,526 2 View

Separate phospholipid monolayer from bilayer?

Hello, I was wondering if it is possible to separate phospholipid monolayered vesicles from bilayered ones? Thanks in advance!

10 February 2017 4,153 7 View

Hemolymph extraction pupa stage?

Hello, Does anyone has experience with extracting hemolymph from the pupa stage of small insects (not Manduca) but for example Drosophila or Spodoptera species? Thank you!

19 October 2016 4,679 2 View