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Questions related to Staining
I should do western blot experiment for detection of protein in liver sample .. i wash tissue with pbs then homogenize it with ripa lysis buffer and PMSF and Na3VO4 and then centrifuge it and...
28 February 2023 6,932 2 View
Hello, What type of statistical test would one use to find a significant difference in antigen expression? (Ex: Surface protein expressed in cancer cells vs. non-cancer cells) would one perform...
28 February 2023 2,142 4 View
Hello All, I have been culturing SH-SY5Y cells and have observed that after plating cells (10,000/well) in 96 well plate, they start clustering by 48 hours (big countable clusters). I am not sure...
28 February 2023 6,349 2 View
I have simple sanger sequencing aligned data with SNPs of about 80 specimens. How can I reconstruct the most likely haplotypes from them?
28 February 2023 1,647 1 View
I use the typical syringe pumps to load organoids (200-300 um in diameter) into my device; however, I don't see them in the chip! I swapped them with beads (similar size) instead and observed that...
27 February 2023 904 2 View
Hello, I'd like to start zebrafish astrocytes, microglia and pericytes culture but I can't really find which are the best conditions and protocol to do that. Also, I may not have the possibility...
24 February 2023 2,083 0 View
My advisor provided me with these protocols. But I am not sure about the protocols which involve so many chemicals. https://zebrafish.org/wiki/health/disease_manual/recipes_and_protocols
23 February 2023 6,240 1 View
I am trying to extract DNA from a starting bacterial culture used for In-vessel composting which came in the hydrogel. I am not sure of the exact type of hydrogel, I tried sonication after...
22 February 2023 3,363 0 View
In the attached paper (by Shankar Balasubramaniam's group) they have used the BG4 antibody to pull down G4 RNA. Does this mean the antibody binds both DNA and RNA, only one with less affinity? Can...
22 February 2023 7,492 3 View
For the same sample these give different values so which one to trust?
21 February 2023 7,573 3 View
I have isolated Naïve CD4 T cells from mouse spleen and tried to polarize them into Th1 cells. For doing that, I have coated a 48-well plate with 1 µg/ml Anti-CD3, 100 µl each well, incubated at...
18 February 2023 3,211 0 View
Both were found on a vessel hull among other macrofouling organisms. Location : Sri Lanka
13 February 2023 6,797 0 View
My lab is looking to purchase a cell counter for tissue culture and I was hoping you guys might have recommendations for ones that you loved (or hated)? At a minimum we’d need one capable of...
12 February 2023 6,862 2 View
We've been trying to transduce THP-1 cells with some P2X7 constructs (Blasticidin resistance gene also present) using Lentivirus and Blasticidin selection. This has worked well in HEK293T cells...
08 February 2023 2,120 2 View
I`m a Ph. D student of mechanical engineering and I need a subject for PHD thesis based on microfluidic systems for testicular sperm extraction. Can anyone guide me and provide me with a topic?
06 February 2023 7,295 0 View
Dear all, I am having problems with the compensation for my 8 color panel. When acquiring my single stain beads, DIVA calculates a spillover of >100% of AlexaFluor700 into APC-Cy7. If I acquire...
06 February 2023 5,057 0 View
I need to transfect cells with a plasmid (GFP). I normally would use a transfection reagent such as lipofectamine or fugene but I only have DharmaFECT 1, which I use for siRNA. Do you think I...
06 February 2023 6,370 4 View
Hi, I fixed chondrocytes in 12-well plate with glutaraldehyde. I want to stain the cells by direct red. Do you have any protocol? Thank you.
05 February 2023 6,881 0 View
Hi, I performed RT-PCR directly on the osteoarthritic cartilages obtained from 2 patients. In both experiments, Ct for TNF-a was found to be around 36, which is unexpected considering the...
01 February 2023 7,248 0 View
I am currently working with HepG2, HepAD38, Huh7 and HepG2-NTCP-K7 cell lines. I want to know what are the correct timeline for those cell lines.
27 January 2023 5,476 3 View
As PMN-MDSCs are considered immature cells - is it possible to culture these? If so, does anyone have culture conditions that preserve the phenotype? For how long? Can they be grown and expanded...
27 January 2023 6,715 0 View
Hello everyone, I'm a phD student doing primary cortical neurons culture for 3 years. I'm doing this protocol from embryonic mice (E16) and following these steps -Dissection in cold HBSS 1X +mg...
26 January 2023 2,431 2 View
We are preparing a transient KD (at the translational level, qPCR is not applicable) and will use targeted proteomics to define the extent of the KD. The KD will be evaluated at 96 hpf (larva...
26 January 2023 8,785 0 View
Dear all, Currently, I am working with RAW 264.7 macrophages. In an experiment, I would like the cells to stop their division and stay at the same number I seed them. Are there some inhibitors...
25 January 2023 9,193 2 View