Polypeptide antigen and antibody related to Tau protein
An antibody and protein technology, applied in the direction of anti-animal/human immunoglobulin, peptide source, animal/human peptide, etc.
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Embodiment 1
[0024] Preparation of polypeptide antigen
[0025] The polypeptide antigen provided by the present invention has the amino acid sequence described in SEQ ID No.1, namely FWSKGDETQGG. Can be synthesized by commercial companies. In this application, the polypeptide antigen was synthesized, purified, and freeze-dried by Zymed Company, and the results are shown in figure 2 , 3 .
Embodiment 2
[0027] Preparation of specific polyclonal antibody (6d) of Tau protein isotype 6d:
[0028] The antigen obtained above was conjugated to KLH [5] After that, immunize animals (rabbits) [6,7] Preparation of antiserum, purification of antiserum to obtain 6d antibody [8,9] , the purified 6d antibody was separated by 10% SDS-PAGE and detected by Coomassi staining ( Figure 4 ) [10] .
[0029] (1) Immunized animals (rabbits) [6,7] Prepare antiserum:
[0030] Add an equal amount of complete Freund's adjuvant to 1 ml of polypeptide antigen solution (1 mg / ml, dissolved in PBS), and emulsify with a double-tube micro-emulsification needle (Thomas Scientific 345OD46). The hair on the back of the rabbit was shaved and disinfected with 70% alcohol. Intradermal injection was performed with a 3ml syringe (24-G) needle at an angle of 30° to the skin, and the injection was divided into 24 points, each injection point was injected with 25 μl, and a total of 3 rabbits were immunized. Six ...
Embodiment 3
[0041] The 6d antibody is used to detect and identify whether Tau protein isoform 6d exists in the form of protein: use 6d as the primary antibody, and detect the expression level and spatiotemporal distribution of endogenous Tau protein isoform 6d in different parts of the brain tissue by Western Blotting; The Tau protein isoform 6d gene was cloned and introduced into COS cells for in vitro expression, and the exogenous Tau protein isoform 6d was detected and identified by Western Blotting. The results showed that A. The new Tau protein isoform 6d did exist in the form of protein ( Figure 5 ,6);
[0042] B. The prepared 6d antibody is a highly specific antibody ( Figure 5 , 6) [11] .
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