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Questions related from Zonghan Gan
Hi! Currently I make fresh Maleimide-PEG-NHS ester solution in PBS to conjugate my protein. However everytime just uls of 1ml made are used. This reagent is not cheap. I saw on MCE that hydrated...
18 February 2025 1,921 2 View
Hi! I have a protein product containing 10% glycerol. Do I need to dialyse to rid off the glycerol first and then add 9 volumes of absolute ethanol, or can I directly add 9 volumes of...
10 February 2025 8,148 3 View
Just bounced on me. Before statistically analysing significant difference, shouldn't we see if data fits normal distribution first? Is 3 replicates enough to testify the hypothesis of normal...
01 August 2024 8,804 13 View
It has been long known that EDC carbodiimide and NHS easter's half life agains hydrolysis are highly related to pH. However, very surprisingly, not yet found a paper giving a table/chart of their...
26 July 2024 9,182 0 View
I noticed that for single-cell mRNA seq, usually N6-12 UMI. But not yet found any paper fully describe how these numbers are calculated. I know roughly a mammalian cell usually has 360,000 mRNA...
26 July 2024 894 0 View
Hi! I notice that reverse transcriptases can be used for both cDNA library construction with template switching (eg. surescript II etc.), and downstream qPCR. To quality-control my library...
16 July 2024 6,376 2 View
Hi! I am staining iNOS and CD163 in my cells so I am looking for a minus control, expressing neither iNOS nor CD163. But the cell familiar in our lab including panc-1 and 293T seems at least...
05 April 2024 8,071 1 View
Hi! I use UV and photoinitiator to gel my PEG-acrylate hydrogel. But I observe that when the temperature is lower (at night during winter), it's very easy to get pre-gelled before molding, even...
02 March 2023 10,092 1 View
Hi! It is known that in wound healing and foreign body reaction, M1 macrophage is responsible for recruiting fibroblast and cause fibrosis. However, studies have also shown that forcibly polarize...
26 October 2022 1,643 0 View
Hi! We gonna to culture some thp-1 derived macrophage on gel. We have some mouse LN111 in stock. Can we just use this to functionalise the gel for adhesion, or if macrophage will be activated...
26 October 2022 1,491 0 View
Hi! It is known that different integrin has impacting macrophage's polarization, eg. ↑integrin b5, or ↓integrin b3, a3, a6 will shift the macrophage to more M2 behaving. Any more brilliant...
25 October 2022 9,603 0 View
Hi! I am looking for a summary/review of overall different integrin isoforms' affinity to different ECM ligand isoforms。 eg. integrin a7b1 has high affinity to Laminin a2, while a3b1 and a6b1 has...
24 October 2022 7,955 2 View
Hi! As in question, the organoid contains artificial hydrogel thus hard to extract single cell solution. Maybe we should start from immuno-staining? Stain surface markers like CD 163, or using...
09 October 2022 9,011 0 View
Is there a review paper in arthritis just like "Hallmarks of cancer" in Cancer research?
25 August 2022 8,926 2 View
Hi! We recently bounce into a concept of integrin internalisation. What we previously involve is mostly integrin clustering and adhesion maturation. Seems both process are triggered by RGD...
15 March 2022 3,468 0 View
Hi! I am trying to input bead into microfluidics. Thus using a syringe with a magnetic stir bar in it. however, beads often aggregate and clog in the plastic hat that connect syringe and needle,...
17 October 2021 729 2 View
Hi! Electrophoresis 2005, 26, 501–510 reported that biotin-streptavidin can be fully, reversibly disassociated in de-ionised water at elevated temperature @ ~70℃. I tried to repeat with 20um...
31 August 2021 10,067 2 View
Hi! Novel sequencing technology (eg. single-cell seq) suggest to use poly dT to capture mRNA. However such technology emphasize to decompose unused single strand poly T primer with exonuclease...
10 August 2021 8,384 1 View
Hi! A special primer has to be used with only 19 bp poly dT (Tm 44.9) instead of 30 bp in original study (Tm 55.2). Will this induce failure of mRNA capture? Will this cause RNA detachment in...
07 August 2021 5,559 3 View
Hello! We are trying to set up a single cell platform using the drop seq technology (Highly Parallel Genome-wide Expression Profiling of Individu... ). However we got some problems Is it better,...
07 August 2021 7,331 0 View
Hi! For some reason we need to adjust the temperature of Reverse transcription when using Superscript II. Is there a activity-temperature curve of Invitrogen Superscript II been profiled in...
04 August 2021 2,198 0 View
Hi! We are trying to use surface functioned microbead for protein conjugation. One major way is to use NHS functioned-bead. The manufactor also recommend the epoxy functioned bead. Both...
02 August 2021 9,177 3 View
Hi! I recently become quite interested in mitochondria research and trying to capture a landscape. What is the most focused phenotypes for mitochondria (including swelling, electron chain...
08 July 2021 2,467 1 View
For some reason, we need to have sodium chloride (NaCl, 0.05-0.1M) in our system. Will this affect the reverse transcriptase and block the SMART reverse transcript? Thanks
18 June 2021 9,998 5 View
Hi! I am quite curious about mitochondrial mechanobiology. How does mechanotransduction (both cell-matrix and cell-cell) affect mitochondial behaviour (mitophagy, mitochondriokinesis & ?) What is...
10 June 2021 5,862 1 View
Hi! I become quite curious about mitochondrial behaviours in tissue. How does mitochondria affect intercellular communication (both cytokine, small molecule, and mechano-signalling? How is the...
10 June 2021 7,145 3 View
Hi! We are using microfluidics to fabricate gel beads. Though they have a central mean, the distribution presents a quite discrete distribution. However for our application we need to strictly...
18 May 2021 7,146 3 View
I am planning to have endonuclease Ⅳ and Bst/Taq work in the same buffer, but definitely in different Temperature, i.e, the system will be passing first 37℃ then 60/95℃. Can I simply put...
14 May 2021 3,298 3 View
I have a DNA sample which in previous step induced Brij 35. can I directly input this sample to PCR reaction? Will existence of Brij 35 interrupt the taq enzyme and the PCR reaction?
13 May 2021 396 3 View
Among many papers a convenient method was used to denature DNA, which is to use the solution of NaOH+Brij. The role of NaOH is easy to understand, but what is the effect of Brij 35...
13 May 2021 3,661 5 View
It is well known that Fpg and Endonuclease IV can cleave AP site in double strand DNA. But can they cleave AP (Apurinic/apyrimidinic) site in single strand DNA?
13 May 2021 3,993 2 View
We aim to cleave our DNA probe from a AP site (apurinic/apyrimidinic site) just rising up the temperature. What is the lowest temperature required to realise the cleavage, 50, 60, 70, 80, or 90...
11 May 2021 179 5 View
Same question in https://forum.image.sc/t/how-to-automatically-measure-transparent-particle-diameter-with-imagej/51754?u=imagejan Hi! I have got some images of transparent gel bead. Because it...
21 April 2021 3,774 3 View
Hi! I am trying to pack one bead per drop using microfluid. I previously assumed that just adjust bead concentration in accordance to how many drop per minute. Yet this route failed. I think I...
04 March 2021 8,418 2 View
HI! I am trying to pack 1 customised hydrogel bead in 1 drop using microfluid, yet the bead ends up outside the drop. The customised bead (diameter 32um) also made from microfluid. The packing...
04 March 2021 7,415 1 View
We are currently researching different surface reversible conjugations between biomolecule and material surface. eg. Streptavidin-biotin can be reversibly disassociate at 70℃ in non-ionic water....
05 February 2021 9,405 0 View
Hi! I noticed that mostly the single cell assay use AMPure beads (magnetic) instead of purifying column. But both function through a DNA affinity coating, just on surface in beads and on membrane...
28 January 2021 9,437 5 View
Hi! We are generating hydrogel beads(50-60um) using microfluidics as single-cell beads. To simply all the experiments (especially the centrifuge), I am looking for a dye. The dye should be stable...
15 January 2021 4,713 8 View
It is known that the Uracil-DNA Glycosylase can remove the basic residue of dU. Thus the DNA chain will further cleave at the point of dU when heating to 95℃ However, is there any method to cleave...
19 December 2020 7,239 3 View
We are designing a probe with hydrogel substrate. Can polyacrylamide survive from 95℃，or is there any other type of hydrogel such as PEG that can survive from this temperature?
19 December 2020 2,901 4 View
Hi! We are aiming to bind streptavidin to polyacrylamide surface. The EDC/NHS method bounced on me as a common method, utilizing the -NH2 of polyacrylamide and -COOH of streptavidin. Is there any...
18 December 2020 8,288 2 View
We are fabricating acrylamide beads as probe substrate. How can we make streptavidin bind to the surface of the acrylamide? Thanks!
10 December 2020 8,445 7 View
Hi! We are designing a genome probe, which need to be cut off the acrydite substrate. Yet the photocleavable spacer is too expensive (＄500 for 50nmol primer, which is just 1 time of assay). Is...
08 December 2020 1,993 2 View
We are designing a nucleotide probe for human gene. We need to then cut off the probe from the substrate. Yet the photocleavable space is tooooo expensive. We are thinking to use an endonuclease....
08 December 2020 6,127 3 View
Hi! We are targeting a gene that has ~100 genotype. We want to design a multiplex primers to capture it from cDNA and doing sequencing afterwards Thus non-specific or degenerate primers are fine...
02 December 2020 9,191 2 View
Hi! After weeks of testing we get control of the drop size, by put oil pump vertical and adjust the oil flow rate. However, we found the drop size becomes crazily deviated when trying to get drop...
29 October 2020 1,294 6 View
Hi! We realise the dripping in our microfluid chip (the + cross). Yet we wonder how can we control our droplet size to ~=70um sphere (the channel is a semicircle in 70um diameter) Can we...
15 October 2020 8,903 8 View
Collagen biomaterial can be heparinised for Growth Factor delivery. Other than the most common EDC/NHS method to heparinise collagen, what are the other methods in vitro? In some study collagen...
22 August 2020 4,583 0 View
It has been long known that collagen matrix can chelate with metal cation like calcium and chromium. That is one way to mix hydroxyapatite with collagen. But for biomaterial, how exactly is the...
19 August 2020 1,330 2 View
It is suggested that as a measurement of the collagen stability, the denature temperature Td is comprised by both the hydrogen bond from triple helix and an entropic part of the fibril. The...
14 August 2020 5,693 1 View
Some studies point that crosslinking increase the inflammatory reaction of collagen[1-3]. while some other studies suggests that Cross-linking also can be introduced to reduce the antigenicity....
11 August 2020 3,477 3 View
I havd found many researches on fabrication of collagen gel always using soluble and insoluble collagen (same-source) respectively. The only difference is the tellopeptide in soluble collagen is...
11 August 2020 4,668 3 View
In investigating collagen I got quite puzzled. Could you please offer me some advice? Here are some of my questions 1. In heterogenous tissue engineering, collagen seems to be potentially the...
10 August 2020 1,010 6 View
Please don't answer because U(T,V) don't have S entropy as argument!!!!!! May I ask a question on thermodynamic? We know that U(V,T) (caloric eq. of state) and S(P,V) (thermodynamic eq of state)...
01 May 2020 7,301 1 View
Hi! we are currently trying to learn of enthesis and especially the in vitro muscle-tendon model of enthesis. Do you know some groups in the area is doing some remarkable and can't be missed work...
24 February 2020 5,123 0 View
Hi! In our project we need to analyze a certain protein secretion by cell. We did a RT-qPCR but the time left is not enough for western blot. We had the immuno-stained fluorescent images of the...
23 November 2019 6,031 3 View
Hi! According to our knowledge, osteopontin (opn) is an extracellular protein. Yet when we discussed the staining image today, some colleagues insisted that opn is intracellullar, surrounding the...
19 November 2019 768 3 View
Hi! We are culturing cells encapsulated in gel matrix and wanna to stain it with alizarin red. Yet the protocol ask to centrifuge the sample to remove the stain while our sample cannot be...
09 September 2019 4,858 0 View
I just read this paper Biomaterials for Integration with 3-D Bioprinting mentioning that "Migration of cells in 3-D can be expedited or minimized depending on the ratios of matrix components...
04 August 2019 7,981 3 View
Hi! We are doing studies about tissue engineering using hybrid scaffold composed of PCL and collagen. However get stuck in RNA extraction for gene expression analysis. Some study suggest using the...
19 July 2019 1,006 3 View
Finding books or other materials systematically explain the structure of protein and how they interact with each other(the forces, the chemical properties). It is even better if it also cover...
11 July 2019 6,534 2 View
Some papers covered that the osteochondral interface, instead of delamination, is gradient, in cell type, cell size, composition and stiffness, and thus a gradient scaffold is required for...
06 June 2019 6,893 0 View
What are the important pathway in TE? I'm a postgraduate just starting working with TE, specifically in our group working with mesenchymal stem cell. In previous study we do cover that how...
02 June 2019 9,571 0 View
Hi! I would like to share a quite interesting paper on microfluidics. A gel shell was made to encapsulate the cell in drop, with conditionable permeability. Though the paper use bacterium, it...
01 January 1970 9,169 0 View
