Manual Protein Extraction Protocol?

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Has someone experience in working with the "Adenosine 5¢-triphosphate (ATP) Bioluminescent Assay Kit" Sigma Aldrich?

Particularly, I want to know the cultured cell ATP assay Protocol, and data analysis (I am working on GBM cell line). Any prompt reply would be highly appreciated Thank you,

09 October 2018 2,430 0 View

Has someone experience in working with the "Adenosine 5¢-triphosphate (ATP) Bioluminescent Assay Kit" Sigma Aldrich?

Particularly, I want to know the cultured cell ATP assay Protocol, and data analysis (I am working on GBM cell line). Thank you,

09 October 2018 6,516 0 View

What is the required concentration of all three (Packaging, Envelop, and Transfer) vectors in Lentivirus Packaging?

I need to package lentivirus to generate gene of interest stable brain tumor cell line so, I want to know what is the required concentration of packaging, envelop and transfer vectors to transfect...

31 December 2017 2,794 2 View

How to interpret JC-10 Staining results

Hello Dear Researchers does anyone have experience to interpret JC-10 staining result for Mitochondria Membrane Potential. Could someone please explain me in detail with images because yet i don't...

11 December 2016 8,419 0 View

What if re-autoclave PBS twice for Cell culture use?

what if re-autoclave PBS twice for Cell culture use? it will work normal or not?

09 October 2016 1,658 0 View

What should be the minimum DMSO concentration to dissolve Temozolomide drug?

Dear researchers i want to treat U-251 cells with Temozolomide (TMZ) so, i want to know that what should be the minimum DMSO concentration to dissolve the Temozolomide and what should be minimum...

09 October 2016 3,123 3 View

Does anyone have exprerience with the situation when Insert and plasmid are not ligating?

I did the digestion of gene X and gene Y with respective enzyme and plasmid concentration and insert concentration and size everything is fine after digestion but after ligation I am not getting...

08 September 2013 8,189 2 View

Not getting desired band of inserts after digesting with respective enzymes?

I want to do subcloning of two genes (gene X and gene Y) in pcDNA 3.1+ expression vector. Gene X is having KpnI and NheI restriction site, gene Y having BamHI and NheI restriction sites....

08 September 2013 3,567 12 View

Is there a problem with my RNA pellet?

Hello, I am currently having problems with RNA extraction. I am using mouse liver (C57BL6J), and I have extracted RNA from mouse liver before. Before this experiment, my final RNA pellets were...

11 August 2024 7,082 3 View

Could it be a cell culture contamination?

Hello everyone! I observed in my culture (htert-RPE1 cells) an orange- red particle at the bottom of the dish. It is visible to the naked eye as a very very small red dot. Could it be a...

09 August 2024 2,824 3 View

RNA Extraction Using Hot Borate Method No Longer Working?

I've been performing RNA extraction on cotton petiole tissue for a few months now using the method described in the following paper, a derivative of the typical hot borate method...

08 August 2024 9,882 2 View

Is there any way to quantify bacterial and fungal cells in their mixed culture?

I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...

07 August 2024 7,535 4 View

Why activated CAR-Jurkat cell could not kill targets?

Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...

06 August 2024 641 2 View

Why does my protein refolded to beta sheet during thermal denaturation analysis?

Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...

06 August 2024 1,989 3 View

How much total RNA concentration to be extracted from sorted plasma cells from bone marrow of C57BL/6 mice for RT-PCR ?

i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...

05 August 2024 8,835 1 View

Low-yield gel extraction problem?

I am having an issue with my gel image where my PCR product is not appearing very bright on the gel. When I perform gel extraction, the A260/280 purity value is very low. I used the Qiagen gel...

05 August 2024 9,798 3 View

Do you have good tips for seaweed tissue preservation in the field for post RNA extraction?

I will be with my students collecting seaweed samples in a marine farm and later we will process this tissue for RNA isolation and further sequencing. Does anyone have tips on how to collect the...

04 August 2024 501 2 View

How to isolate lymphocytes from mouse spleen?

I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...

04 August 2024 9,913 7 View