Penta·His Alexa Fluor 647 Conjugate

Protocol tips

Upstream tips	Protocol tips	Downstream tips

Publication protocol

Library screening: Recombinant human His-tagged CEACAM5 extracellular domain as well as recombinant human His-tagged HER2 extracellular domain were purchased at R&D systems. For library screening, respective library cells were grown overnight in SD-Trp medium at 30 °C and 200 rpm. Afterwards, cells were transferred to SG-Trp medium at approximately 107 cells/mL followed by incubation at 20 °C for 2 days. In general, antigen binding was detected by application of Penta-His Alexa Fluor 647 Conjugate antibody (Qiagen, 1:20 diluted in PBS). Fab display on the yeast surface was visualized using light chain specific goat F(ab’)2 anti-human kappa R-phycoerythrin (SouthernBiotech, 1:20 diluted in PBS). Labeling steps were performed with 107 cells/20 µL on ice.
Labeling of cells for FACS analysis or sorting was conducted by two consecutive washing steps of library candidates with PBS followed by incubation with the respective antigen at a concentration of 1 µM. After incubation on ice for approx. 30 min an additional washing step was performed, followed by staining with Penta-His Alexa Fluor 647 Conjugate antibody as well as light chain specific goat F(ab’)2 anti-human kappa R-phycoerythrin. Finally, cells were washed with PBS. FACS-sorting rounds were either performed on a Sony SH800 cell sorter (Sony) or on a MoFlo Legacy cell sorter (Beckman Coulter).



Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Protein tag Detection of His-tagged proteins using Penta·His Alexa Fluor 647 Conjugate from Qiagen.

Manufacturer protocol

Download the product protocol from Qiagen for Penta·His Alexa Fluor 647 Conjugate below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Protein tag Detection of His-tagged proteins using Penta·His Alexa Fluor 647 Conjugate from Qiagen. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.