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.|
|Reactivity:||tdTomato (also recognizes mCherry)|
|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 Chan's Lab, Northwestern University, submitted February 23, 2015.
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