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EM Immunogold Labeling Protocol - P(4)

时间:2006-01-22 15:57来源:ihcworld.com 作者:bioguider 点击: 1181次

1% Sodium Borohydrite Solution in 0.1M PB:

To prepare 100 ml,

Sodium borohydrite -------------- 1 g

0.1M PB ---------------------------- 100 ml

Stir to dissolve.

 

Blocking Buffer (1% BSA, 3% NGS, 0.04% Triton in PBS):

To prepare 100 ml,

Triton X-100 ----------------------- 0.04 ml

BSA ---------------------------------- 1 g

Normal goat serum -------------- 3 ml

PBS ---------------------------------- 100 ml

Stir to dissolve.

 

Washing Buffer (0.8% BSA, 0.1% Fish Gelatin in PBS).

To prepare 100 ml,

BSA ---------------------------------- 0.8 g

Fish gelatin ------------------------ 0.1 ml

PBS --------------------------------- 100 ml

 

Secondary Antibody:

1nm gold conjugated secondary antibody from Amersham. Dilute the antibody 1:50 in washing buffer.

                                                                                                      

Silver Enhancement Solution:

Kit from Amersham and prepare solution according to the instruction provided.

 

Embed-812:

Embed-812 Kit from EMS containing Embed-812, DDSA, NMA, and DMP-30. Make Embed-812 according to instruction provided with the kit.

 

1:1 Mixture of Embed-812 & Propylene Oxide:

To prepare 20 ml,

Embed-812 ------------------------- 10 ml

Propylene oxide ------------------- 10 ml

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