HRP storage, SPDP and TMB suppliers.

BSS169 at BANGOR.AC.UK BSS169 at BANGOR.AC.UK
Fri Dec 23 10:01:35 EST 1994


>From:  CBS%UK.AC.NSFNET-RELAY::CN.AC.IHEP.BEPC2::WANGYQ 21-DEC-1994 >09:41:18.20
>Subj:  enzyme storage

>Dear internetter:
>Does there anyone who can tell me a method of long time storage of enzyme
>especially Horse Radish Peroxidase, at work concentration?
>Please tell me where I can buy SPDP and TMB, or does anyone can supply
>small amount?
>Thank you in advance.
>My address is: 9-702, An Wai Dong He Yan
>              Dong Cheng District
>               Beijing 100011, P.R.China
>Tel/Fax: 86-1-4252418
>e-mail: Wangyq at bepc2.ihep.ac.cn


1. You can store the HRP in 2-10 mM sodium phosphate buffer, pH 5.5-6.5 at 
-70C after degassing, (if  necessary, you can raise the pH by adding Na2HPO4 
before conjugation). As a general rule, repeated freezing-thawing should be 
avoided, so, distribute you HRP solution into small aliquots.  I don't know 
what your buffer and its working concentration and pH are, but be aware of 
possible precipitation of the HRP at high concentration, due to its neutral 
or slightly basic pI (depending on what isozymes it mainly contains), and 
possible inactivation of HRP by polystyrene (e.g. microplates), NaN3, and O2. 

2. From PIERCE company, you can buy SPDP, Cat. No.s: 21557, 21657, 21650, 
21651. and TMB, Cat. No.s: 34015, 34016, 34017, 34021. The address of  
PIERCE's distributor in China is:
Sino American Biotechnology Co.
126 Dao Bei Road,
LUO YANG,
TEL. 1-203-5607
FAX. 1-202-0354.

You also can buy SPDP and TMB from CalBiochem. Company. Cat. No.s: 573112 
(SPDP); 613544 (water soluble TMB) and 613545, 613548 (insoluble TMB). 
A distributor nearest to you is in Hong Kong, 
SM Biochemical Company Ltd.
18-B capital Building,
6-10, Sun Wui Road,
Causeway Bay, 
Hong Kong 
Tel. 852-5765038
Fax. 852-890-1979.

The water soluble TMB is for ELISA and insoluble TMB is for blots detection 
and immunohistochemical analysis.  

It seems that you are going to conjugate HRP to something by linker SPDP 
and visibilize by TMB. There are many (maybe too many) linkers and 
chromogens/substrates available technically, you may have to try 
somethings else later.   

Good luck!

Dr Dan Wu

School of Biol. Sci.,
University of Wales,  
Bangor LL57 2UW
UK

fax. 44-248-371644
e-mail: bss169 at bangor.ac.uk
    



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