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I am in need of dihydrorhodamine (DHR) respiratory burst assay to evaluate phagocytes function in patients suspected for chronic granulomatous disease. Is there any practical protocol that you...
07 March 2021 4,468 3 View
Dear all, We would like to check the purity of a GST-tagged recombinant protein by SDS-PAGE. We purified the protein using Baculovirus expression system. We run the reducing SDS-PAGE (GST was...
19 December 2016 1,925 9 View
Dear all, I would like to conduct beta-Galactosidase assay. The protocol requires a permeabilization step using the recipe below: for one sample, we need: 100 mM dibasic sodium phosphate...
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Dear all, I would like to ask for your advice with culturing culture Eol-1 cells (eosinophilic cell line). I resuscitated cells from frozen stock and suspended in RMPI 1640+ 10% FBS + 1% PS. After...
02 May 2016 6,850 8 View
Dear all, I would like to purify lung mononuclear cells from BALB/c mouse and detect CD4+ T cells by FACS. At first, I tried to isolate cell suspension by collagenase digestion and to proceed to...
07 December 2015 5,305 7 View
I used OneComp eBeads (eBioscience) to set up the compensation for PE & PE-Cy7. However, in the single PE staining tube, I couldn't achieve two distinct populations with similar mean values. I...
07 October 2015 3,446 6 View
Dear all, I'm currently trying to detect the signals of intracellular LC3 in isolated eosinophil from human blood. After being purified (the purity is ~ 99%) and treated, eosinophils are...
13 April 2015 2,280 3 View
I need to gate eosinophil on FACS by using double markers CDw125 (surface markers) and EPX (intracellular proteins). Here I already had PE antiCDw125, but for EPX, I need to do indirect staining....
03 February 2015 9,144 7 View
Hello all, I am trying to purify basophil from human whole blood. I previously used 6% Dextran sedimentation to isolate red blood cells and others, and I collected basophil contaminated with...
02 September 2014 8,185 11 View
To perform the total cell counts in BAL fluid recovered from an OVA-induced mouse asthma model, we usually dilute BAL fluid 1:1 with Trypan Blue, then count 4 corner squares of hemocytometer. One...
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