Nano gold coupled antibody / protein Kit (20nm, 1mg/100 μ g/500 μ G coupling amount) preparation

Nano gold conjugated antibody / Protein kits can be tested by antibodies / The primary amine group on the protein （ Such as lysine ）, Simple 、 Quickly realize antibody / Covalent coupling of protein and nano gold . Coupled at 30 In minutes , The coupling efficiency can reach 70% about . This is a general-purpose kit , Suitable for antibodies of various molecular weights / protein , It is also applicable to polypeptides 、 Enzymes and other molecules . Now let's share with you Nano gold conjugated antibody / Protein kit （20nm,1mg/100μg/500μg Coupling amount ） Storage and steps of , Watch it with Xiaobian ！
Kit composition and storage

Store the kit in -20⁰C. coupling agent 、 The reaction buffer can be stored in +4℃ or -20℃

The experimental steps

Ø Restore all reagents to room temperature .

Ø take 500 μL Add reaction buffer solution 1 In gold nanotube , Get the dispersion of nano gold ; take 1 The tube coupling reagent is dissolved in 200 μL In pure water .

Ø take 100 μg antibody / Add the protein into the nano gold buffer solution in the previous step , In this mixed solution , Then add 100 μL Coupling reagent , After mixing, continue at room temperature 30 About minutes .（ Be careful ： Coupling reagent needs to be prepared and used now ！）

Ø After the reaction, use 9500 rpm, centrifugal 20 minute , Wash with pure water 2~3 Times to remove the unreacted antibody / protein .

Ø Conjugate antibody storage ： Labeled antibody / Post protein , We suggest storing the conjugate in +4°C, Do not store conjugates in -20°C. The overall stability of the conjugate will be determined by the antibody / The stability of the protein itself determines , Because it will degrade first . If your antibody / Protein stability , The conjugate will also be stable .

Ø Measure the concentration of the conjugate ： Nano gold (20nm) Maximum absorbance (Absmax) by 528 nm. In order to determine the effective concentration of nano gold conjugate , We recommend diluting the sample to a suitable range for your equipment , Use UV - Measured by visible spectrophotometer Absmax.

matters needing attention

Ø This kit is compatible with amine free buffer （PBS, MES, MOPS, HEPES） and < 50% glycerol . If the sample antibody / Protein contains the following components , May interfere with the sample antibody / Coupling of protein and nano gold .

Whether the buffer composition is compatible with amine free buffer （ Such as ：PBS, MES, MOPS, HEPES） yes (pH 6.5-8.5) Glycerol is (＜ 50%)Gelatin ( gelatin ) no Tris No glycine no carboxylic acid molecule (EDTA、 Citric acid, etc ) Whether it contains primary amino group or sulfhydryl molecule ( Amino acids, 、 Ethanolamine 、 Mercaptoethanol or DTT etc. ) No serum protein No .

Ø If the above interference components exist , The interfering components can be removed first by means of ultrafiltration tube or dialysis , Then use this kit to transfer the antibody / Protein is coupled with nano gold .

Ø Best conjugated antibody / Protein content ： Conjugated antibody / The amount of protein will affect the antibody on the surface of each nano gold particle / Number of protein molecules , Best conjugated antibody / The amount of protein depends on your application needs . therefore , You may need to couple different amounts of antibodies / protein , Combined with the application test optimization, the best conjugated antibody / Protein content .

Ruixi YWX.2022.7.28