This antibody was raised against a recombinant protein and recognizes tdTomato.
The tdTomato protein was derived from a monomeric mutant of DsRed, a red fluorescent protein from so-called disc corals of the genus Discosoma. tdTomato is an important tool in biotechnology and cell biology as a spectrally distinct companion or substitute for the green fluorescent protein (GFP; from Aequorea jellyfish).
From a laboratory at The Scripps Research Institute.
|Antigen:||Raised against a recombinant protein and recognizes tdTomato.|
|Purification Method:||Protein G|
|Buffer:||0.1M Sodium Phosphate, pH 7.4, 0.15M NaCl, 0.05% (w/v) Sodium Azide|
|Tested Applications:||WB (1:1000), ELISA, IF, IHC (1:100-500), IP (1ug antibody to 100ug total protein)|
|Storage:||4C, -20C for long term storage|
Immunohistochemistry using Anti-tdTomato [16D7] Antibody in a transgenetic mouse, weakly expressing Cre-driven tdTomato.
The Anti-tdTomato [16D7] Antibod with Alexa-555 secondary was used to visualize Cre+ cells for tdTomato expression. The protocol used is as follows: (1) Perfuse mouse with ~ 25 ml ice-cold normal ringer solution with pH 7.4. (2) Perfuse mouse with ~ 50 ml ice-cold fixative consisting of 2% PFA and 15% picric acid with pH = 7.4. (3) After brain extraction, post fix for one hour with 2% PFA + 15% picric acid. (4) Quickly wash brain 3x with liberal amount of PBS, thoroughly removing all fixative. (5) Slice in ice-cold ringer and section at 60 µm. Wash section 3x with 0.01M PBS, 15 each time. (6) Block for 1 hours with 10% normal goat serum INGS), 3% Bovine serum albumin (BSA), and 0.1% Trixton-X in 0.01M PBS. (7) Incubate primary tdTomato Ab at 1:500 1:250 for 24 hours at 4°C on a shaker. (8) Extract primary solution without washing. (9) Incubate for 2 hours in room temperature secondary (Alexa-555) at 1:500 in the following solution made in 0.01M PBS: 1% NGS + 0.01% Triton-X. (10) Wash sections 3x, 15 each time. (11) Mount section on microscope slide and cover slip using anti-fading preservative (e.g. Vectashield or Fluoromount-G). Image courtesy of Kerafast customer Vivian Hernandez from Dr. C. Savio Chans Lab, Northwestern University, submitted February 23, 2015.
If you publish research with this product, please let us know so we can cite your paper.