Application
ELISA
Sample
Human Cell (Supernatants of carcinoma cells)
Specification
Supernatants of carcinoma cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Type
Indirect
Other product details
Incubation time
2 hour(s) and 0 minute(s) · Temperature: 20°C · Diluent: PBS
Concentration
1 µg/ml
Secondary antibody
Dilution
1/5000
Name
Non-Abcam antibody was used: Santa Cruz goat anti rabbit-HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: HRP polymer
Host species: Goat
Clonality: Polyclonal
Conjugation: HRP polymer
Additional data
Additional Notes
500 000 cells were incubated with 1 ml serumfree medium for 24h, supernatants were harvested and 100 µl of supernatant, or diltutions of recombinant SDF were applied to a Nunc maxisorp 96 well plate for 2h at 20°C. Blocking with 1%BSA/0.05 Tween in PBS, washing with PBS. Antibody application as listed above, detection with TMB substrate (Pierce) and 0.5M H2SO4 as stop solution, measured at 450 nm. Standard curve was good, and detection limit is about 1 ng/well (10 ng/ml), could probably be also further reduced.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
DR. Kirsten Hattermann
Verified customer
Submitted May 15 2013