Debaprasad Koner

16 Questions 45 Answers 0 Followers

Questions related from Debaprasad Koner

How many cells I need to make protein sample from primary hepatocyte culture for WB analysis?

I need to check protein expression in primary hepatocytes after some treatment through WB analysis. I am facing problem regarding amount of protein in cell homogenate. The protein concentration...

08 August 2019 6,162 9 View

How to synthesis PbO nanoparticles by chemical process?

I want to make PbO nanoparticle through chemical process. Can anyone provide me the protocol. Thanks in advance

09 September 2018 6,257 2 View

How to raise primary antibody against my protein of interest?

I want to raise primary antibody against my protein of interest in my lab. I need help to make it. I have full length mRNA sequence of my gene. Can you provide me any protocol to raise primary...

09 September 2018 9,038 6 View

How to check tissue specific distribution of mRNA using Real-time PCR?

I just want to check the expression of a particular mRNA throughout the different tissues (Liver, Kidney, Brain, Muscle, Gills, Intestine, Ovary, Skin, Spleen etc.) in a control/untreated fish. If...

02 February 2018 5,366 7 View

Why only BSA and Skimmed milk proteins are used for blocking in Western blotting?

Why antibodies (primary and secondary) do not cross react with these protein? What chemistry is there to choose these proteins as blocking reagent?

01 January 2018 3,600 7 View

Do the dilution factor of antibody (primary and secondary) should be same for endogenous control and target protein?

The same dilution factor of antibody doesn't fit for the endogenous and target protein. I am working on fish model and the primary antibody which I am using for catalase (CAT) protein needs...

10 October 2017 3,584 3 View

Have anyone used Amplex® Red Hydrogen Peroxide/Peroxidase Assay Kit for H2O2 quantification from cell using Qubit?

Dear Researchers, I am using Amplex® Red Hydrogen Peroxide/Peroxidase Assay Kit (Thermo Fischer) for H2O2 quantification from cell. There is an option for Qubit but, there is no clear protocol for...

11 November 2016 6,998 2 View

Is it recommended to use 0.5X SYBR Green Master Mix?

I use StepOnePlus Real-Time PCR System for relative gene expression studies. I follow SYBR Green chemistry. Is it advisable to use 0.5X SYBR Green Master Mix instead of 1X? Thanks in advance.

08 August 2016 5,374 8 View

What is the optimum concentration of Nitric Oxide (NO) in brain tissue of Catfish (Clarias batrachus)?

I have quantified NO in different tissues of catfish (Clarias batrachus) using Griess reagent. I am getting very high amount of NO (>300 nmoles/ gm wet wt). I have followed the steps...

08 August 2016 4,745 1 View

What is the difference in the different parts of fish kidney in terms of functional anatomy ?

Dear all, How do kidney parts (head, mid and tail) of fish differs in their functional aspect ? Thanks in advance.

08 August 2016 5,442 0 View

What should be the unit of Nitric Oxide in tissue?

I have estimated Nitric Oxide from tissue homogenate by using griess reagent. From the standard curve, I got a concentration about 12 µM. I have used 100 µl of tissue homogenate. The protein...

01 January 2016 7,523 5 View

Should I use DTT (Dithiothreitol) or any reducing agent in homogenizing buffer for enzyme assay?

I want to assay the enzyme activity of different antioxidant enzymes. Should I use any reducing agent in homogenizing buffer? I have gone through some protocols for assay and the reducing agents...

01 January 2016 5,373 15 View

What is the significance of molar extinction coefficient in enzyme kinetics assay?

What does it actually imply?

01 January 2016 6,547 3 View

Why am I not getting any result in WB from the primary hepatocyte culture lysate?

From the primary hepatocyte culture of C. batrachus, I am getting protein concentration in the range of 7-10 µg/µl in Bradford assay. However, in western blotting, I use to load 50-75 µg total...

12 December 2015 9,730 5 View

What is the best way to make ZnO nanoparticle suspension in water to assay its toxicity in fish?

I want to check ZnO NP toxicity in fish. Before to give treatment, I used to sonicate the ZnO NP in water for 10 minutes at 30 kHz. What should be the best way to make ZnO NP suspension for giving...

12 December 2015 7,960 5 View

How to prove the expression of only one single isoform of a certain enzyme?

In our fish (C. batrachus), from the transcriptome data we have found several isoforms of the enzyme GST. However, following ZnO nanoparticle treatment fish shows a expression of only a single...

12 December 2015 7,231 2 View