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Generation of phospho-cPLA2(pcPLA2T268) antibody

SP Sangita Paul
SF Saman Fatihi
SS Srishti Sharma
RK Rintu Kutum
RF Raymond Fields
HP Harish C. Pant
LT Lipi Thukral
BB Binukumar BK
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The custom-made pcPLA2T268 antibody was generated by the GenScript Antibody Group [the cPLA2_268-phospho-antigen 1-(LLL{pTHR}PQKVKRYVESC); lot 779114-1, GenScript; the cPLA2_268- non-phospho-antigen 2 (LLLTPQKVKRYVESC), lot: 779115-1, GenScript; Peptide (L>OT: 779114-1)-KLH conjugate, ImmunoGen; host stain, New Zealand Rabbit].

The cPLA2_268-phospho-antigen 1-(LLL{pTHR}PQKVKRYVESC) with keyhole limpet hemocyanin (KLH) conjugate was injected into the New Zealand rabbit for immunization. The serum was isolated and subjected to cPLA2 non-phosphopeptide affinity column for subtraction. The flow through was further subjected to the phosphopeptide affinity column for affinity purification. The antibodies binding to the phosphopeptide column was eluted and characterized further through indirect ELISA. For antibody characterization, the phosphopeptides and nonphosphopeptides were coated with 4 μg/ml, 100 μl/well concentration with PBS, pH 7.4, as coating buffer. The secondary antibody used in the ELISA was anti-rabbit IgG monoclonal antibody (HRP conjugate; catalog #AO1827, GeneScript).

Copyright and License information:  ©2022 Paul et al.
This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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