...type and the protein of interest. You may need to try a variety of fixation methods to find the best conditions... Protocols Immunocytochemistry Immunocytochemistry You may also like... Antibody Plasmid...Antibody Purification Protocol Introduction Immunocytochemistry is a technique that uses antibodies to detect...

...your primer length and increase the Tm if you are trying to clone from genomic DNA, a cDNA library, or by...to conduct 2 ligations for each plasmid you are trying to create, with varying ratios of recipient plasmid...colonies on your recipient plasmid alone plate, you can try ligating the recipient plasmid alone in the presence... that your transformation worked. You could also try varying the amount of recipient plasmid to insert...

...to conduct 2 ligations for each plasmid you are trying to create, with varying ratios of recipient plasmid...colonies on your recipient plasmid alone plate, you can try ligating the recipient plasmid alone in the presence...you are plating on the appropriate antibiotic and try varying the amount of recipient plasmid to insert...

...glycerol stock before you begin your plasmid mini-prep. Try not to freeze/thaw your glycerol stock too many times...

... that you cannot perform your duties. Be sure to try out different sizes to know your fit. It is best ...

...instead of incubating at -20°C for 1 hour, you can try incubating at -80°C overnight. Centrifuge for 20 ...

...protect the UV box from being cut by the razor blade. Try to get as little excess gel around the band as possible...

...growth after overnight incubation. What went wrong? Try growing the culture for more time. Some bacterial...following into a 500 mL glass bottle: 4 g NaCl 4 g Tryptone 2 g Yeast Extract and dH 2 O to 400 mL Note: If...

...site (MCS), but does not cut your insert, you could try using two different enzymes that have compatible ...

...do controls. See Tips and FAQ below for details. Try different vector to insert ratios to optimize the...

... a microscope’s eyepiece can take some practice. Try to keep both eyes open and adjust the position of...

... a sample of possible dilutions. You may want to try higher/lower dilutions depending on your downstream...

... purposes. What do I do if my PCR isn't working? Try adding 1µl of 25mM MgCl 2 and/or 1µl DMSO to each...

...location of the protein of interest. You may need to try a variety of lysis buffers to find the best for your...Collection Antibody Blog Posts Antibody Guide Immunocytochemistry Protocol Recombinant Antibody Purification...

...microfuge tube and vortex. Pro-Tip You may need to try a few different concentrations of antigen to determine...101: The Four ELISAs and When to Use Them Immunocytochemistry Protocol Recombinant Antibody Purification...

...the closed system of a crowded plate or culture. Try restricting growth to the bare minimum by growing...

...Opti-MEM, Thermo Fisher 31985-070 0.05% Trypsin/EDTA (e.g. TrypLE, Thermo Fisher, 12605010) 1X PBS pH 7.4...Aspirate PBS and add 2 mL of 0.05% Trypsin/EDTA. Wait ~2 min. Neutralize trypsin by adding 10 mL of DMEM Complete...new vial of cells after 30 passages. Procedure Trypsinize and resuspend the HEK293T cells from 2 x T-175...cells in 1 CS2. Return to incubator for 48 h. Trypsinize and resuspend cells from the CS2. Cells should...60 mL PBS. Aspirate PBS and add 35 mL of 0.05% Trypsin/EDTA. Wait ~2-3 minutes for cells to detach. Gently...until no live cells can be seen when stained with Trypan Blue. Pellet cell debris by centrifugation at 3900...

...affect the attachment of adherent cells) Trypsin EDTA (TrypLE, Thermo Fisher 12605-010) 6-well tissue ... Isolate cells from the stable cell pool by trypsinization and break up any cell clumps into individual...expanded but before they become over-confluent, trypsinize the cells and expand them to larger culture dishes...the independent monoclonal lines. Pro-Tip When trypsinizing cells from the 96-well plate, use a microscope...detached from the plate and can be isolated. When trypsinized, detached cells will exhibit a rounded morphology...

...containing the trypan blue. Pipette 10 times to mix. Load 10 µL of the cell suspension/trypan blue mix into....45 µm PES filtering system, 500 mL, VWR 430770 Trypan Blue, Thermo Fisher T10282 Valproic acid sodium...flask 5–10 times before sampling. Transfer 10 µL of trypan blue into a clean microcentrifuge tube. Vortex ...

...affect the attachment of adherent cells) Trypsin EDTA (TrypLE, Thermo Fisher, 12605010) Ethanol, VWR, ...each well. Detach cells by incubating with 200 µL TrypLE for 1–2 min. Resuspend cells in 500 µL DMEM complete...