[Preparation and preliminary application of rabbit polyclonal antibodies against intracellular peptides of human platelet glycoprotein GPIbalpha].
OBJECTIVE
To prepare rabbit polyclonal antibodies against intracellular peptides of human platelet glycoprotein GPIbalpha.


METHODS
Two peptides corresponding to human platelet GPIbalpha C-terminus were synthesized and purified by high-performance liquid chromatography (HPLC). The peptides were cross-linked with keyhole limpet hemocyanin (KLH). Two New Zealand white rabbits were immunized with conjugated peptides for 3 times. The polyclonal antibodies were purified by Ammonium Sulfate Precipitation and identified by dot blotting and ELISA. GPIbalpha intracellular peptides phosphorylation was tested with these polyclonal antibodies by ELISA.


RESULTS
The titers of the two polyclonal antibodies against the GPIbalpha C-terminus peptides were 1:32 000 and 1:64 000 respectively and both of these antibodies reacted with purified GPIbalpha.


CONCLUSIONS
Two rabbit polyclonal antibodies against C-terminal peptides of human platelet GPIbalpha have been prepared successfully, providing a way for the preparation of these kinds of antibody. Both phosphorylation and dephosphorylation states exist in the intracellular peptide of human platelets.
Yuan
Yan-Hong
YH
Hematology and Cell Biology Laboratory, School of Biological Science and Medical Engineering, Beihang University, Beijing 100083, China.
Kang
Jia-Rui
JR
Yan
Rong
R
Cheng
Hong
H
Zhuang
Feng-Yuan
FY
Wang
Yue-Dan
YD
Dai
Ke-Sheng
KS
D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D006801:Humans; D008562:Membrane Glycoproteins / Q000276:immunology*; D010768:Phosphoserine / Q000737:chemistry; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000276:immunology; D011817:Rabbits
D000906:Antibodies; D008562:Membrane Glycoproteins; D019038:Platelet Glycoprotein GPIb-IX Complex; C094990:adhesion receptor; D010768:Phosphoserine