Direct ELISA analysis of Phosphotyrosine, Phosphoserine, and Phosphothreonine was performed by coating wells of a 96-well plate with 100ul per well of Phosphotyrosine, Phosphoserine or Phosphothreonine-BSA conjugate diluted in carbonate/bicarbonate buffer (Product # 28382) at a concentration of 5ug/ml overnight at 4°C. Wells of the plate were washed, blocked with Superblock blocking buffer (Product # 37515), and incubated with 100ul per well of a rabbit anti-phosphotyrosine,serine and threonine polyclonal antibody (Product # 61-8300) at a starting concentration of 1ug/ml and serially diluting 2-fold to a concentration of 8ng/ml for 1 hour at 37C. The plate was washed, then incubated with 100ul per well of an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31430) at a dilution of 1:8000 for 30 minutes at 37C. Detection was performed using 1-Step Ultra TMB substrate (Product # 34028) for 5 minutes at room temperature in the dark. The reaction was stopped with 0.16M sulfuric acid, and absorbances were read on a spectrophotometer at 450-550nm.
|Tested species reactivity||Chemical|
|Published species reactivity||Non-human primate, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Phosphothreonine conjugated to protein carriers|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||1 µg/ml|
|Immunoprecipitation (IP)||5-10 ug|
|Western Blot (WB)||1.0 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This antibody detects pS, pT and pY amino acids with very low cross reactivity to their non-phosphorylated variants
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
In vitro effects of recombinant otoconin 90 upon calcite crystal growth. Significance of tertiary structure.
61-8300 was used in western blot to assess the effects of recombinant Otoconin 90 in vitro
|Lu W,Zhou D,Freeman JJ,Thalmann I,Ornitz DM,Thalmann R||Hearing research (268:172)||2010|
|Non-human primate||Not Cited||Conserved SNH domain of the proto-oncoprotein SYT interacts with components of the human chromatin remodelling complexes, while the QPGY repeat domain forms homo-oligomers.||Perani M,Ingram CJ,Cooper CS,Garrett MD,Goodwin GH||Oncogene (22:8156)||2003|
|Mouse||Not Cited||IkappaB kinase-independent IkappaBalpha degradation pathway: functional NF-kappaB activity and implications for cancer therapy.||Tergaonkar V,Bottero V,Ikawa M,Li Q,Verma IM||Molecular and cellular biology (23:8070)||2003|
|Mouse||Not Cited||Prostaglandin levels in stimulated macrophages are controlled by phospholipase A2-activating protein and by activation of phospholipase C and D.||Ribardo DA,Crowe SE,Kuhl KR,Peterson JW,Chopra AK||The Journal of biological chemistry (276:5467)||2001|
||Prostaglandin levels in stimulated macrophages are controlled by phospholipase A2-activating protein and by activation of phospholipase C and D.||Ribardo DA,Crowe SE,Kuhl KR,Peterson JW,Chopra AK||The Journal of biological chemistry (276:5467)||2001|