08 August 2011 2 8K Report

I want to express a human receptor on HEK cells. Is there anywhere, like a database or something, where I can find out if the HEK-293 cells have endogenous expression of the receptor or do I have to do qPCR for that?

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I am working with ELISA and use HRP conjugate as detection antibody and TMB substrate. All of the protocols I have used so far the TMB substrate reaction has been stopped by adding an acidic...

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Why does my standard curve not generate expected concentrations in the back calculation?

I am setting up a direct ELISA and have a standard curve which I fit to a 5-PL curve. When I try to calculate the concentrations of the points in the standard curve using the obtained 5-PL curve I...

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Is it possible to efficiently conduct E coli cell lysis for protein purification with ultrasound bath?

I want to lyze my E. coli for protein purification but I only have an ultrasound bath, not a sonicator with a probe. Does anyone have any experience of this? Any tips or protocols that can help me.

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Can anyone help with "Shadows" of bands on non-reducing a SDS gel?

I have run a non-reducing gel with samples of a few antibodies. The samples are supposed to be pure, i.e. only containing the antibody of interest. I observe some bands below the main band. Does...

07 August 2014 9,815 7 View

What is a good software for curve fitting?

I am looking for a good software for fitting curves and would like to hear what software other people recommend. I want to be able to fit 4- and 5- parameter logistic curves and linear regression....

03 April 2014 6,133 45 View

Difference between FreeStyle 293 (Gibco) and Pro293-DM (Lonza)?

I am fairly new to cell cultivation and I am wondering about the main differences between the FreeStyle 293 media from Gibco and the Pro293-DM from Lonza. As I understand it both are serum and...

04 May 2013 9,474 2 View

Optimal cell (HEK-293) density for transient transfection and IgG expression

I am wondering what your opinion is on what cell density is optimal for expression of antibodies by transient transfection of suspension adapted HEK-293 cells in batch culture using PEI as...

01 February 2013 9,611 2 View

How to conjugate dGDP to protein?

I want to conjugate cyclic GDP to a protein to use in immunization for antibody production. Can anyone help me with a way in which I can do this conjugation?

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Why fix cells before ELISA?

I have a basic question I wish I knew the answer to. In several protocols for doing ELISA to study internalization of receptors or internalization of antibodies I see that they fix the cells...

11 December 2012 7,417 6 View

Protein concentration using Amicon Ultra spin column, adsorption on membrane?

I am trying to concentrate a protein using Amicon Ultra centrifugal device. I have done this several times before without any problems. This time I had a larger volume of my sample so I had to...

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