Application
ELISA
Sample
Mouse Cell (NB41A3 cells)
Specification
NB41A3 cells
Type
Sandwich (Capture)
Blocking step
3% BSA (Sigma) + 1% FBS in PBST (0.05% Tween-20) as blocking agent for 40 minute(s) · Concentration: 0.1% · Temperature: 25°C
Other product details
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: PBS
Dilution
1/1000
Secondary antibody
Name
Non-Abcam antibody was used: Phosphotyrosine [pY] Monoclonal Antibody, Mouse (P
Host species: Mouse
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Host species: Mouse
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/500
Additional data
Additional Notes
This antibody was used as a capture antibody for endogenous mouse RET in a sandwich ELISA.The ELISA plate was coated with 100 uL of 1:1000 dilution of ab134100 overnight at 4 C and blocked for 45 min with 300 uL of blocking buffer (3% BSA, 1% FBS in PBST). Mouse neuroblastoma NB41A3 cells stably transfected with GFRa3 were treated with ART or assay buffer for 10 min at 37C and lysed. The lysates were transfered to the ELISA plate and incubated for 2 hours at room temperature with shaking. The plate was washed with PBST and incubated with anti-phosphoTyrosine antibody conjugated to HRP for 1 hour at room temperature and washed. 100 uL of TMB/well was added to the plate and incubed until blue color developed (5 min). The reaction was quenched with 100 uL of 1M sulfuric acid per well. Absorbance was measured at 450 nM.
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提交于 Mar 03 2015