[Preparation, identification, and analysis on tissue chips of polyclonal anti-peptide antibody to chemokine-like factor 1].
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.


CONCLUSIONS
The anti-peptide antibody can specifically recognize CKLFs and may be a useful reagent for the detection of CKLF1.
Shi
Shuang
S
Department of Immunology of Basic Medical Sciences College, Center for Human Disease Genomics, Peking University, Beijing 100083, China.
Zhang
Ying-mei
YM
Qiu
Xiao-yan
XY
Zhu
Xiao-hui
XH
Liu
Ya-nan
YN
Gan
Qi-ni
QN
Yang
Tian
T
Song
Quan-sheng
QS
Ma
Da-long
DL
Han
Wen-ling
WL
D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D018925:Chemokines / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D006801:Humans; D062792:MARVEL Domain-Containing Proteins; D020411:Oligonucleotide Array Sequence Analysis; D010446:Peptide Fragments / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D011817:Rabbits; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology
D000906:Antibodies; C432676:CKLF protein, human; D018925:Chemokines; D062792:MARVEL Domain-Containing Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins