IF-FISH experiments were performed as described before (Takai et al. 2003 (link)). The primary antibodies used were 53BP1 (1:1000; rabbit polyclonal; Novus Biologicals 100-304A) and Myc (1:1000; Cell Signaling 9B11). The secondary antibodies used were raised against rabbit or mouse and conjugated with Alexa 488 (Invitrogen) or Rhodamine Red-X (Invitrogen), and diluted 1:1000. Telomere FISH was performed using Alexa 488 or Cy3-OO-TelC-labeled PNA probes (PNA Bio, Inc.). Streptavidin Alexa 488 ( 1:2000 dilution; Invitrogen S32354, ) was used for detection of biotinylated proteins by IF.
Alexa 488
Manufactured by PNA Bio
Alexa 488 is a fluorescent dye that can be used for labeling and detection purposes in various biological applications. It has an excitation maximum at 495 nm and an emission maximum at 519 nm, making it suitable for use with common fluorescence detection systems.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using alexa 488
1
Visualizing DNA Damage Repair Foci
2
Telomere Length Measurement by FlowFISH
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TL was measured by flowFISH following the detailed protocol in Baerlocher et al. (24 (link)), except that peripheral blood mononuclear cell were extracted by density gradient separation before preserving in freezing media. Red cells were lysed at the time of thawing (RBC Lysis Buffer; eBioscience). Cow thymus was obtained from a local butcher and fixed thymocytes were mixed with each sample (24 (link)). A peptide nucleic acid (PNA) labeled probe containing the sequence (CCCTAA)3 was used for the hybridization (Alexa 488; PNA Bio). Each sample was run in triplicate in addition to a no probe to account for background (24 (link)). The average lymphocyte and granulocyte TL were then calculated. A reference sample with known TL, obtained from a large leukapheresis product, was used to normalize fluorescence values on each plate. For this sample, the leukocyte TL was measured by Southern blot (1 (link)), as well as by flowFISH at a company (Repeat Diagnostics). Intra- and interassay CV was calculated by dividing the SD (σ) by the mean (µ).
3
Immunofluorescence and FISH Experiments
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
IF-FISH experiments were performed as described before (Takai et al. 2003) . The primary antibodies used were 53BP1 (100-304A, rabbit polyclonal; Novus Biologicals; 1:1000), Myc (9B11, Cell Signaling; 1:1000). The secondary antibodies used were raised against rabbit or mouse and conjugated with Alexa 488 (Invitrogen) or Rhodamine Red-X (Invitrogen), and diluted 1:1000. Telomere FISH was performed using Alexa 488 or Cy3-OO-TelC labeled PNA probes (PNA Bio Inc.). Streptavidin-Alexa 488 (Invitrogen #S32354, 1:2000 dilution) was used for detection of biotinylated proteins by IF.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!