Preparation And Identification Of Punctin-2 Polyclonal Antibody

Punctin-2,also known as ADAMTSL-3(a disintegrin-like and metalloprotease domain with thrombospondin type I motifs-like-3),is a member of the subfamily ADAMTSL in the ADAMTS protein superfamily.ADAMTS protease family has catalytic activity of metalloproteinases and binding activity of the auxiliary domain,ADAMTSL members have only provide binding activity of the auxiliary domain,there is no domain of metalloproteinases.Punctin-2 is a secretory glycoprotein in the extracellular matrix(ECM),and widely exists in human tissues.The study found that the expression of punctin-2 in bone is closely related to height.At the same time,the mutation of punctin-2 found in colon cancer may be related to the occurrence of cancer.In C.elegans motoneurons,madd-4 causes motoneurons to be excited and inhibited by affecting the postsynaptic localization of acetylcholine receptor(AChR)and GABA receptor(GABARs).We know the function of madd4 in C.elegans motoneurons,and then we want to explore the function of punctin-2 in mammalian nervous system.At present,there is no effective antibody to punctin-2,so we prepare the antibody of punctin-2 to study the role of punctin-2 in the brain.Through the analysis of the amino acid sequence of mouse Punctin-2,three antigenic peptides were synthesized and coupled to immunize New Zealand white rabbits to obtain serum and purified antibodies.Affinity purification method was used to purify the serum to obtain antibody.ELISA,WB and Co-IP experiments were used to detect and identify the antiserum and purified antibody.