AI Article Synopsis
- The study aimed to create a polyclonal anti-peptide antibody against CKLF1 and utilize it for examining CKLF1 expression and function.
- The researchers selected a 16 amino acid sequence from the C-terminal end of CKLF1 and immunized rabbits with this peptide to generate the antibody.
- Results showed that the antibody effectively recognized CKLF1 in various tissues and had significant staining differences between normal and cancerous rectal tissues, indicating its potential as a tool for CKLF1 detection.
Article Abstract
Objective: To prepare the polyclonal anti-peptide antibody against chemokine-like factor1 (CKLF1) and apply it to the expression and functional studies of CKLF1.
Methods: CKLF1 was analyzed with bioinformatics methods. The 16 amino acids sequence peptide was selected from CKLF1 C terminal end. Antibody was raised by immunizing rabbits with the peptide conjugated to keyhole limpet hemocyanin (KLH).
Results: A high titer polycolonal antibody was obtained from the rabbit against the peptide. ELISA analysis proved that the titer of rabbit serum against anti-peptide of CKLF1 was up to 10(-4). Western blot analysis revealed that it could react not only with recombinant CKLF1 expressed in a cell-Free Protein Biosynthesis System and Drosophila S2 cells, but also recognize the endogenous CKLFs in the tissue array. Positive staining was detected in the normal bronchial cartilage, gastric mucosa, and gastric smooth muscle tissues. Normal rectum and well-differentiated rectal carcinoma showed strong positive staining, but the poor-differentiated rectal carcinoma samples revealed negative staining.
Conclusion: The anti-peptide antibody can specifically recognize CKLFs and may be a useful reagent for the detection of CKLF1.
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