Martin Forde

12 Questions 61 Answers 0 Followers

Questions related from Martin Forde

Can I use blocking peptide as positive control in Western blot?

I'm just curious if blocking peptides can be used in this fashion? Does the conjugation of the blocking peptide with the primary antibody stop the immunoreactivity with say a flouro-tagged...

01 January 2016 5,633 5 View

Contaminated taqman probe?

i think my taqman primer / probe is contaminated. is there any way to decontaminate? e.g., uv light, etc.

09 September 2015 226 2 View

MDA-MB-231 advice?

I'm looking to do a time course realtime-glo vitality assay on MDA-MB-231 in a 96-well plate. I plan on doing 4 different seeding concentrations to assess linearity of signal, however there is...

06 June 2015 1,852 4 View

How can I perform SiRNA concentration scaling? by nM or pmol per num of cell / surface area?

I'm looking to scale up my siRNA reverse transfections from a 6-well to 384-well plate for adherent SH-SY5Y. Should I be scaling the siRNA concentration by surface area in the well, or by the...

01 January 2015 4,828 6 View

How can I explain great siRNA transfection efficiency but no knockdown with validated positive controls?

I'm performing reverse transfection on SH-SY5Y using Viromer Green, and I get wonderful transfection efficiency (90+%). See picture (RFP labeled siRNA fluorescence overlapped on phase contrast...

11 November 2014 7,547 7 View

Can anybody recommend how much Blasticidin should be used for selection of SH-SY5Y?

We are electroporating a pcDNA6.2 plasmid contain miR RNAi constructs into SH-SY5Y. I plan on selecting these cells with blasticidin, and the literature suggests closer to the 10ug/ml mark for...

10 October 2014 6,510 3 View

Is anyone familiar with electroporation?

I thoroughly pipette my cells prior to electroporating so they should form a monolayer when plated. Though every time I electroporate, they clump. I'm not entirely sure why. When I passage, they...

09 September 2014 6,382 3 View

Shsy5y immunostaine troubleshooting

cells plated

08 August 2014 2,251 0 View

What is a good number of SH-SY5Y to freeze down for long term storage between 1-12mo?

What is a good number of SH-SY5Y to freeze down for long term storage between 1-12mo? Ideally, I'd like to initially plate in something larger like a t75 or t150, though I'm not sure if it is...

07 July 2014 4,806 2 View

What is the best method to carry out SH-SY5Y differentiation and transfection?

How do people typically go about electroporating differentiated SH-SY5Y? I was thinking that you would have to electroporate them first then recover in media supplemented with retinoic acid. Would...

07 July 2014 1,769 1 View

Can anyone share their knowledge on cell penetrating proteins?

Has anyone had any experience using CPPs to transfect small plasmids or siRNA into primary human neuron? I'm just curious to see if you would share your experiences. I'm going to be testing with...

07 July 2014 1,461 2 View

Can somebody help with combinatorial shRNA from concatamers?

Would it be feasible to treat multiple independent domains as a concatamer? E.g., Forward 5' = Sense1 -- Sense2 -- hairpin -- AntiSense2 -- AntiSense1 -- term Reverse 5' = term -- S1 -- S2 --...

04 April 2014 2,516 0 View