Does any of you have protocol for insect cell (Sf9) expression protocol?

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Protein purification from Sf9 insect cells

Hi, I am trying to purify the chimeric fusion proteins containing NGF and various functional domains of Fibronectin. Although lower molecular weight proteins were secreted into the conditioned...

01 February 2017 8,084 6 View

NGF Characterization assays on PC12 Cells?

can any one suggest what potential experiments could be performed to test the NGF activity on PC12 cells except Neurite outgrowth, migration & proliferation.

03 April 2016 9,182 3 View

Any suggestions on the cell lysis buffer for his tag protein purification?

Hi Researchers, Following to my previous questions, few of my chimeras are unable to secrete into media as expected but present in cells. Current plan is to purify from cells and I tried once with...

04 May 2015 5,106 4 View

It is mandatory to remove imidazole after recom protein purification for cell based functional assays?

Hello researchers, I'm working on chimeric fusion proteins (NGF:FN) growth factors and ECM protein combination. I have expressed this protein in insect cell system and purified on Cobalt resin...

02 March 2015 2,015 13 View

Does anyone have set hands in purifying the his tagged recom protein (secretory) NGF?

I'm working on Novel protein therapeutics for peripheral nerve regeneration. My protein NGF (Nerve Growth Factor) was expressed in Sf9 cells and secretory protein was unable to purify under native...

09 October 2014 356 4 View

Why Do TDS and EC Increase with Larger Wastewater Volumes, While BOD and COD Decrease?

I have carried out MFC experiments on three different volumes, 50, 500 and 1000 mL of wastewater. Results after MFC treatment shows that TDS and EC are more in larger volumes of water i.e. TDS and...

09 August 2024 9,621 0 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

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

Can I use a HisTRAP column for affinity chromatography?

I'm working on selecting antibodies against a recombinant protein that has a His-tag. My idea is to first bind the recombinant protein to a HisTRAP column and then use this column for an affinity...

07 August 2024 505 3 View

How to normalize and take the significance of the MTT OD values with 3 replicates for the same cell-line?

Hi, I have a question about normalizing the MTT OD values for doing the statistical analysis. So, if we have 3 different plates and we call them 3 different replicates, so, first we would...

07 August 2024 8,106 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 can I improve the quality of lamb milk replacer for machine feeding?

Hello everyone, I am researching ways to enhance the quality of lamb milk replacer for machine feeding. I would appreciate any insights or recommendations on the following: Manual vs. Machine...

06 August 2024 6,227 2 View

How can i do multivariate Time Series forecast using MLP, ANFIS and LSTM?

I need the python code to forecast what crop production will be in the next decade considering climate and crop production variables as seen in the attached.csv file.

05 August 2024 2,977 3 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