Can anyone please help me with a protocol for in-vitro phosphorylation using CAMK2α ?

07 June 2022 0 6K Report

I want to phosphorylate my protein using CAMK2α. I ordered CAMK2α Kinase Enzyme System from promega (https://www.promega.de/en/products/cell-signaling/kinase-assays-and-kinase-biology/camk2-alpha-kinase-enzyme-system/?catNum=V4018).

However, I can not find a protocol of howmuch kinase/substrate/buffer to use. If anyone has used this system, could you please share your protocol?

Badges
Science topic

More Satabdee Mohapatra's questions See All

I need MD simulation in 500 ns for some of my test compounds with PBSA, GBSA and PCA. If a researcher will help, we will provide authorship?

For my research, I need MD simulation of 500 ns with PCA. Please reply if you have facility. We will provide corresponding authorship. The article will be published in a high IF journal. My email-...

26 November 2023 6,926 0 View

How to increase the solubility of chlorophyll extracted from plants for bioimaging studies?

Please provide information related to any coating that can be done or any composites that can be synthesized.

09 October 2023 9,003 2 View

As we all know that Si 2p peak can be deconvoluted into two peaks such as Si 2p1/2 and Si 2p3/2. Which peak has higher binding energy and why ? ?

In the few literature, it was mentioned that the binding energy of Si 2p1/2 was higher than Si 2p3/2, while some authors made the just opposite statement about these peaks. I am so confused about...

22 June 2023 1,135 2 View

What is the JCPDS No and CCDC no for Ag-MOF-5?

Ag-MOF-5 synthesized from Terepthalic acid, Silver Nitrate and DMF. Need to know Chemical formula, Molecular weight and molecular structure of Ag-MOF-5. Thank you

21 May 2023 1,623 0 View

Can a composite show different dual phases (space group) in room temperature XRD and room temperature Neutron diffraction Reitveld refinement?

If possible kindly suggest some references.

05 February 2023 3,682 3 View

How to isolate fibroblast cells from hamster?

I isolated fibroblast cells from hamster by collagenase A digestion method. I used DMEM with extra L-glutamine as culture media. After 24hr there is no attachment of cells. The flask was coated...

28 November 2022 4,815 1 View

Which is the best technique- FACS or MACS for separation of dead cell from live cell population by using PI and SYTO9 fluorescence dye?

I want to separate dead cells from a bacterial population after treatment of any drug or antimicrobial agent. Which technique will be more efficient FACS or MACS if I use PI and SYTO9 dye ? Is...

15 August 2022 4,008 3 View

How to separate my metal microcage synthesised in ethylene glycol and water solvent ??

I tried to pellet it down at a maximum of 18K rpm, but it did not work. I heated the reaction mixture up to 200 C temp to evaporate the Ethylene Glycol(Including water) and to get the metal...

21 June 2022 6,740 0 View

Is it possible to cryopreserve the Neonatal primary cardiomyocyte?

Neonatal cardiomyocyte is the sensitive primary cell line. Can a researcher cryopreserve the cell line for future experiment?

30 December 2021 4,160 3 View

What is the reason behind negative Z' value in EIS measurement?

I did the electrochemical impedance measurement using a three-electrode system and I am getting a negative Z' value at a higher frequency (1MHZ) in my Nyquist plots. can anyone explain what could...

16 November 2021 3,012 3 View

How to analyze multiple phosphorilation sites?

Hello, I am currently analyzing some phosphoproteomics data, but I have peptides with multiple phosphorylation sites or phosphorylations together with carbamidomethylation or oxidation. How can I...

04 August 2024 8,432 3 View

When you express a protein, why do we express not only the domain we want, but also the protein around it?

I want to express STK4, and I've searched the paper for reference. When I check the protein kinase domain sequence for that kinase on Uniprot, it's 30-281, but the paper expresses the protein...

20 July 2024 4,951 1 View

Phosphorylation sites prediction?

Hello, Does anybody have idea about any software or database that can give information about the possible phospho sites located in a protein? I am trying PhosIDP but its not working or loading...

15 July 2024 1,940 3 View

How to detect phosphorylation on Western Blots?

Having trouble detecting phosphorylated HSL. Any tips for how to prevent de-phosphorylation events or detect phosphorylation?

15 July 2024 2,055 3 View

Medium replacement interfering with cell culture blot results?

We utilize an immortal endothelial cell line and various agonists to observe their effects on Src phosphorylation. However, our protocols seem to induce high basal levels of Src phosphorylation....

17 June 2024 1,649 0 View

PVDF or nitrocellulose to visualize phosphorylated proteins?

Which membrane material is the best for capturing and visualizing phosphorylated proteins in a western blot? Or is there no difference?

05 June 2024 9,502 2 View

What are the options for testing the phosphorylation of HP1a?

I have used CSII to phosphorylate HP1a, in vitro. Now, to test this phosphorylation, I have options such as using an anti-serine phospho antibody or mass spectrometry. Has anyone ever tested an...

08 April 2024 1,952 2 View

Why IBMX but not forskolin during adipocyte differentiiation?

During preadiopcyte in vitro differentiation, cocktail stimuli is needed. But why IBMX but not forskolin? Both of them can increase the intracellular level of cAMP and lead to PKA pathway...

28 February 2024 8,146 0 View

Can Thymidylate kinase (TMK) targetting antibiotics have any effect on human TMK?

Hello, I'm working on an antibiotic against MRSA by targeting thymidylate kinase (TMK); however, I do not know if this antibiotic will have any negative or positive effects on human thymidylate...

24 February 2024 889 0 View

Recommendations for phospho-serine and/or phospho-threonine antibody?

Can anyone recommend a "good" general phospho-serine and/or phospho-threonine antibody for use after immunoprecipitation in a western blot? There are a lot of expensive offers online with...

22 February 2024 1,080 0 View