Thanks for the inquiry. The volume of the 2x loading buffer depends on the loading amount into the well. For example, if you would like to use 30μg/well as the loading amount, it should take out 6μl as a sample from 5ug/ul whole-cell/tissue extract or the recombinant protein. And 2x loading buffer should use 6μl for each sample (2x sample buffer: sample volume is 1:1). Then you can follow Step10 on the video to finish all steps for the sample preparation. We hope it helps. Please feel free to contact us if you have any questions. www.genetex.com/contact
@@Genetex_Antibodies actually I'm getting different band density for beta actin.... I am loading 20ug of protein concentration for 20ul well.... Can you suggest something
@@itssushmita687 Thanks for sharing the feedback with us. As the result you got, we think the concentration of samples has some problems. You can consider redetermining the concentration of samples by using a BSA standard curve. We hope this information will be of use. Please do not hesitate to contact us if you have any questions. The contact address is support@genetex.com.
@@itssushmita687 It is possible that your treatment to the cells is affecting the house keeping gene expression which actually signifies that you need ton check your choice of House Keeping Gene. It is possible that Beta-Actin is not suitable for your cells or treatment
![Western Blot - Semi-Dry transfer [Video from GeneTex]](http://i.ytimg.com/vi/krFKPwhiHJE/mqdefault.jpg)
Nice and simple! Love it
Thanks, GeneTex appreciate your support!
I was looking for like this video for a long. Thank you for sharing with us =)
so informative
thank you
It's helping me a lot. Thank you
How much 2x loading buffer to be added for 5ug/ul of protein?? Please tell me
Thanks for the inquiry.
The volume of the 2x loading buffer depends on the loading amount into the well.
For example, if you would like to use 30μg/well as the loading amount, it should take out 6μl as a sample from 5ug/ul whole-cell/tissue extract or the recombinant protein. And 2x loading buffer should use 6μl for each sample (2x sample buffer: sample volume is 1:1). Then you can follow Step10 on the video to finish all steps for the sample preparation.
We hope it helps.
Please feel free to contact us if you have any questions. www.genetex.com/contact
@@Genetex_Antibodies thank you so much
@@Genetex_Antibodies actually I'm getting different band density for beta actin.... I am loading 20ug of protein concentration for 20ul well.... Can you suggest something
@@itssushmita687 Thanks for sharing the feedback with us.
As the result you got, we think the concentration of samples has some problems. You can consider redetermining the concentration of samples by using a BSA standard curve.
We hope this information will be of use.
Please do not hesitate to contact us if you have any questions. The contact address is support@genetex.com.
@@itssushmita687 It is possible that your treatment to the cells is affecting the house keeping gene expression which actually signifies that you need ton check your choice of House Keeping Gene. It is possible that Beta-Actin is not suitable for your cells or treatment