The detection of several types of cancer is possible by using monoclonal antibody (MAb). Studies have shown that different types of cancer have related specific abnormal proteins, which are identified by MAb. IBMR3 is a MAb which was raised against synthetic peptides that are associated with the selected amino acid sequence of human interleukin-4 receptors. Aims: The aim of this study was to evaluate the effectiveness of specific monoclonal IBMR3 antibodies expression in the cancer cell lines of 3T3 and HT29, in order to recognize specific antigen and make evaluation, using immunoblotting technique which is widely use in this filed. Methods: Protein extracts were extracted from these cancer cell lines and used for western bloat. The Immunoblotting were consequently subjected to densitometry analysis using bio imaging machine. This bio imaging process will facilitate to measure the molecular weights, peak height and raw volume of the protein bands for the 3T3 and HT29 cancer cell lines, which helps in diagnosis of any pathogenic antigen. Results: The bands obtained from bio imaging were exposed on the PVDF membrane. In the 3T3 bio imaging process revealed four bands and molecular weight were 299.58, 87.90, 41.67 and 23.54 KDa. However, bio imaging results for HT29 revealed also four bands with molecular weights: 90.11, 41.31, 23.87 and 20.86 KDa. The results of Peak height densitometry for IBMR3 antigen bands for 3T3 were: 1856.985, 551.769, 394.164, and 216.185. However HT29 peak height results were: 281.544, 101.711.202.668, 757.213. The raw volume (amount of protein bands of IBMR3 Ag for 3T3) were: 1460168.75, 206078.47, 161406.89, and 219583.16. However HT29 raw volume results were: 110197.11, 76106.84, 98632.59, and 221395.34. Negative protein staining for 3T3 and HT29 cancer cell line bands were done by using mouse IgM serotype. IgM serotype was not indicated that means no specific antigens for negative control IgM. Conclusion: The bio imaging revealed ........