Is it harmful to breed for Cre Homozygosity initially?

Hello, I am currently using a Cas9 knock in system that will be expressed in the liver. To do so STOP codon fl/fl Cas9 KI mice were bred with Albumin-Cre mice to generate heterozgous pups, that...

24 February 2021 5,759 3 View

Does anyone has a protocol for transfecting AML12 cells?

I am optimizing a CRISPR/CAS9 system in vitro before using it in vivo. I would like to transfect AML12 cells with my pAAV-sgRNA-vector to validate editing efficiency before generating virus.

26 January 2021 8,733 2 View

The impact of Methylisothiazolinone on haemoglobin quantification?

Hello everyone, I am a researcher working on a project aiming to develop an assay for Haemoglobin quantification. In our assay, prior to HGB quantification we dilute the sample in a dilution...

20 November 2020 7,316 2 View

Ansys Workbench Project view shows "Solved" but mechanical shows "Solve Required". How to Fix this without resolving ?

Hello everyone I am running a thermo-mechanical simulation on a Linux Cluster. In my Project view all the component systems seem to have solved. There is a file.rst file and and solver.out...

14 October 2020 2,631 3 View

Standard guitar tuning has two distinguished vectors (0 5 5 5 4 5) and (0 5 10 15 19 24). What is required to show they are tangent-cotangent bundles?

The pitch value set in music and guitar group in tablature are connected by adjunction of tangent-cotangent bundles. Tuning g is the tangent gradient to the flow of pitch on guitar. It determines...

04 April 2020 2,681 15 View

Can you ask for user input on pymol?

I want to write a script that asks the user to enter the PDB file so that the script can be entered (it prompts the user to enter it), the user enters the 4 character code, then the rest of the...

23 February 2020 1,184 1 View

Why cani I not find Constantino Manuel Torresd on Research Gate Search Enginge?

I just read a new paper posted here by Constantino Manuel Torres. However when I put his name in the search engine here, it says there is now one here with that name? I wanted to send him a...

13 July 2019 2,496 3 View

How do I design primers specific to transfected plasmid and not genomic DNA?

I recently introduced a Lenti-Puro Guide vector containing a specific sgRNA sequence into a cell line. I would like to sequence the ddCas9-sgRNA cell lines to confirm that each sgRNA is accurate.

04 February 2019 6,681 3 View

How to make culture media more basic (not too basic) without a pH meter or strips?

Hi, A friend of mine suggested that the reaction that makes Salmonella colonies appear black on XLD or TSI agar might show an increased rate at a slightly lower pH. I was thinking of adding a bit...

24 July 2018 2,916 7 View

Does anyone has protocol for p53 IHC in frozen sections?

I need to detect p53 in murine E15.5 embryos that overexpress this protein. The embryos were collected and fixed overnight in 10% Neutral Buffered Formalin. Then, I cryopreserved these embryos in...

23 May 2018 2,592 2 View

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View

What is the most suitable resuspension buffer for TCA/acetone precipitated protein?

I need to extract protein from fermented carbon sources for Bradford assay. Most researchers experiencing insolubility of pellet in resuspension buffer. Please assist me to select most suitable...

26 February 2021 7,129 3 View

EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View

Standard curve shifting over time. How do I stabilise the ELISA?

24 February 2021 1,365 3 View

What is the proper in vitro culturing method to get information about the activation of innate immune cells by VLPs?

Dear All, I am interested in studying the response of immune cells against virus-like particles (VLPs). I used the PBMC in vitro assay and treat it with the VLP using PHA as a positive ctrl while...

16 February 2021 1,337 2 View

Why does flow deformation occur at the test line and control line on nitrocellulose membranes (LFIA)?

I use 1% BSA in 10 mM phosphate buffer (pH 7.4) solution when striping my nitrocellulose membranes with antigens and antibodies. However, I always observe deformed flow when my sample or running...

16 February 2021 4,224 3 View

Can the viability assays (ex: MTT/CCK-8) measure cell proliferation? or are they solely viability assays?

I want to conduct a proliferation assay to test a chemotherapy combination on a cancer cell line. At first, I thought about the MTT assay. But now, all sources are saying that these do not measure...

14 February 2021 3,142 7 View

GST assay in microplate?

I'd like to know if anyone can provide me with a protocol for traditional GST enzymatic assay (with GSH and CDNB as substrates) in a 96 well microplate, for cell line and/or C.elegans...

11 February 2021 1,328 3 View

What lysis buffer is compatible with Bradford assay?

Hello! I am planning to use Bradford assay to measure protein content in PBMCs lysates. The problem is that Bradford assay doesn't seem to be compatible with Tris-based buffers or SDS. Are there...

10 February 2021 1,797 5 View

For PC-3 cell line. How long before irradiation do I have to synchronize my cells to G0/G1?

I am trying to do a clonogenic assay for PC-3 after irradiation. I have to synchronize my cells by serum starvation but there's a optimal time I have to do that before irradiation. been looking in...

09 February 2021 9,480 1 View