Damian D Guerra

9 Questions 18 Answers 0 Followers

Questions related from Damian D Guerra

Oops! I added the wrong secondary to my Western blot! Can I still obtain a clear signal without stripping the blot?

-I want to detect differences in protein expression via LiCoR near-infrared secondaries. -I probe with a RABBIT primary antibody, against my favorite protein. -I add the *wrong* secondary--Goat...

12 December 2017 4,148 4 View

Immunofluorescence Dual Antibody Labeling Question: can I use a secondary antibody made in the same organism as one of my primary antibodies?

Say that I am going to perform an immunofluorescence experiment in which I want to see the localization and abundance of two proteins, X and Y. I have the following antibodies: Primary: Mouse anti...

09 September 2016 2,932 10 View

Are Qiagen RNeasy kits suitable for extraction of yeast RNA for downstream RNAseq?

I was wondering if anyone could advise me on RNA quality. I used a Qiagen RNeasy kit to isolate RNA from yeast and an Agilent Bioanalyzer to check the quality of the RNA (see attached). Yeast...

06 June 2016 9,274 0 View

Storage recommendations for peptides?

I have immunoprecipitated my protein of interest, along with putative interacting proteins. This mixture was resolved by SDS-PAGE, and proteins were in-gel digested by trypsin. Resulting peptides...

06 June 2016 5,548 4 View

Any suggestions about an Li-COR for plants?

I am trying to perform a western blot against a protein expressed in plants. The antibody is well-characterized, the protein target is abundant, and I have previously visualized the protein in...

12 December 2015 7,340 5 View

How long after a floral dip will Agrobacterium tumifaciens continue to be capable of transforming the infected plant?

For example, if I dip an Arabidopsis plant today, will flowers that appear one week from today still be capable of being transformed? This is assuming "normative" growth conditions (ie, 20 degrees...

01 January 2015 3,713 14 View

Is it necessary to sequence the entirety of my insert after an LR reaction if I know that the sequence of my entry clone is good?

Expression from a Gateway vector requires two cloning steps: First, a PCR amplification product is cloned into an entry vector (pENTR or pDONR) using BP clonase. Secondly, the entry construct is...

09 September 2014 1,887 9 View

Is it likely that a 25% (w/v) sodium dodecylsulfate (SDS) solution will be a source of contamination?

My colleague and I are working with immortalized mammalian smooth muscle cells. We are re-using a modified 96-well plate. Thus far, this plate has shown no signs of contamination. However, to...

01 January 1970 8,277 4 View

Akt phospho antibody--suggestions?

I am attempting to detect differences in Akt phosphorylation at Serine 473 with Cell Signaling Technology product #9271 (Rabbit anti-phospho-Akt). Typical lysis conditions and standard SDS-PAGE,...

01 January 1970 6,305 3 View