Figure 4: Human apolipoprotein L1 antibody. A) A peptide corresponding to amino acids 125-139 of human apolipoprotein L1 or ApoL1 (highlighted in gray) was selected for generating a new antibody. Input parameters for AbDesigner: Input Type = Gene Symbol-Human, Input = APOL1, Peptide Length = 15, Epitope Length = 7. B) Immunoblotting of lysates from CHO cells transfected with pCMV>GFP and/or increasing amounts of pCMV>ApoL1-GFP plasmid DNA as shown was probed with the K5 rabbit anti-ApoL1 antibody. The 62.5 kDa band that intensifies with increasing plasmid amount is of the expected molecular weight for ApoL1-GFP protein. C) The membrane shown in (B) was blotted with mouse anti-GFP (700 nm channel) and the K5 rabbit anti-ApoL1 antibody (800 nm channel). The yellow band in the merged image indicates colocalization between the 700 nm and 800 nm channels at 62.5 kDa. D) Competitive blocking of the K5 antigen peptide was used in increasing ratio to the K5 antibody. E) Representative image from indirect immunofluorescence microscopy of HeLa cells transfected with 5 ng/μl pCMV>ApoL1- His6 or empty vector control and stained for ApoL1 using the K5 anti-ApoL1 antibody. No signal was observed in cells transfected with empty vector, whereas the pCMV>ApoL1 transfected cells displayed a robust cytoplasmic signal, particularly in a perinuclear location. The experiment was repeated four times with similar results.