[Production of camel polyclonal antibody against vascular endothelial growth factor receptor-2 and performance evaluation in ELISA test]

In molecular approach, serum of camel contains a unique type of antibodies devoid of light chains since the light chain is missing, the heavy-chain antibodies should bind their antigen by one single domain, the variable domain of the heavy immunoglobulin chain. Vascular endothelial growth factor receptor-2 [VEGFR-2] is one of the important proteins in angiogenesis which over-expressed in many types of tumors. Two male dromedaries [Camelus dromedarius] were immunized against VEGFR-2. ELISA was used to evaluate the immunization process. Camel immune sera were fractionated with protein A and G affinity chromatography. Heavy chain antibodies tested for its specific reactivity in cell-based ELISA in recognition of VEGFR-2 on the cell surface of three cell lines; 293/KDR, HUVECs and A431. In ELISA test, the camel sera in 1/12800 dilution had the acceptable results. Furthermore, cell-based ELISA demonstrated that the polyclonal heavy chain antibodies bind to VEGFR-2 in 293/KDR and HUVECs cell lines. Single chain polyclonal antibodies against VEGFR-2 can be used for detection of soluble form of this protein by ELISA, and also flowcytometry, western blotting and immunohistochemistry

allele and genotype frequencies of bovine pituitary - specific transcription factor and leptin genes in Iranian cattle and buffalo populations using PCR-RFLP

The use of polymorphic markers in breeding programmes could make selection more accurate and efficient. A total of 324 individuals from six Iranian cattle populations [Sarabi, Golpayegani, Sistani, Taleshi, Mazandarani, Dashtiyari], F1 Golpayegani Brown Swiss and Iranian buffalo populations were genotyped for the Pit-1 HinfI and leptin Sau3AI polymorphisms by the polymerase chain reaction and restriction fragment length polymorphism [PCR-RFLP]. The genotype and gene frequencies for each breed were determined and shown to be quite variable among the breeds. The highest frequencies of allele B for the leptin gene and allele A for the Pit-1 gene were found in Dashtiyari and Sistani cattle, respectively. According to our results, the highest AB genotype frequencies were found in the Taleshi and F1 Golpayegani x Brown Swiss cross for the leptin and Pit-1 genes, respectively. These allele frequencies were comparable to previously published data on exotic breeds. The highest and lowest heterozygosities were found in Taleshi and Dashtiyari cattle for the leptin gene and in F1 Golpayegani x Brown Swiss cross and Sistani cattle for the Pit-1 gene, respectively. These values indicated the presence of low variation for these genes in the studied populations. The possible association between molecular polymorphisms within these candidate genes and economic traits for the studied populations should be further investigated