Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Protocols and Methods Forum
Register Search Today's Posts Mark Forums Read

Protocols and Methods Forum Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.

for estimation of acid phosphatase

for estimation of acid phosphatase - Protocols and Methods Forum

for estimation of acid phosphatase - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.

LinkBack Thread Tools Display Modes
Old 09-26-2008, 11:11 AM
satheesh kumar
Posts: n/a
Default for estimation of acid phosphatase


Im satheesh kumar, i plan to estimate the acid phosphatase activity in
coffea. so anybody wil send me some our good suggestion to me...
Reply With Quote
Old 09-26-2008, 09:07 PM
Posts: n/a
Default for estimation of acid phosphatase


cheapest way should be pNPP (p-nitrophenyl phosphate) as substrate. If
you prefer higher sensitivity and/or prefer fluorescent detection, you
might choose MUPF (methyl umbelliferyl phosphate) or ELF phosphate,
just to mention some possibilities. If you want to use
chemiluminescence, check CSPD/CPD-star substrates. pH is adjusted with
appropriate buffer in all cases.

Best reagrds,


On Sep 26, 1:11*pm, "satheesh kumar" <[Only registered users see links. ]> wrote:

Reply With Quote
Old 03-03-2014, 08:36 AM
Zagami's Avatar
Ph.D Doctorate
Points: 4,670, Level: 46 Points: 4,670, Level: 46 Points: 4,670, Level: 46
Activity: 0% Activity: 0% Activity: 0%
Join Date: Sep 2009
Location: Tabaka Mission Hospital - Kiisi -Kenya & Italy Sicily
Posts: 175
Thanks: 2
Thanked 28 Times in 24 Posts
Default Re: for estimation of acid phosphatase

Acid phosphatase (AcP) hydrolyze disodium phenylphosphate in acidic medium (pH 4.9) to form phosphate and phenol. In fixed time, reaction is, by sodium arsenate, stopped and phenol liberated is condensed by 4-aminoanipyrine in alkaline (pH 7.9 ± 0.1 or 10.0) medium and in presence of potassium ferricyanide in a red complex that showed a absorption peak about 500 – 520 nm. This method, by me modified, is applicable in the phenol concentration range of 1 μg/L to 250 μg/L with a sensitivity of μg/L.
Substrate buffer (citrate buffer 0.2 M at pH 4.9 containing 10 mM of disodium phenylphosphate) : dissolve in 200 ml dH2O, 21.0 g di citric acid monohydrate, add 188 ml of 1 M NaOH and dilute to 500 ml with dH2O. Adjust the pH to 4.9, if necessary, with molar NaOH or molar HC1, then dissolve 10 mM of disodium phenylphosphate, 05 g/L NaN3 and keep at 4 °C, stable one year in opaque plastic bottle.
Blocking reagent : 30 mM 4-aminoantipyrine, 120 mM sodium arsenate dibasic heptahydrate (Fluka cod. 71625) in NH4OH/NH4Cl buffer at pH 10 ± 0.2 (dissolve 67.5 g of ammonium chloride in 570 ml of ammonium hydroxyde, and add freshly boiled and cooled water to make 1 liter), and 0.5 g/L NaN3. Keep at 4 °C, stable one year in opaque plastic bottle.
Color reagent : 75 mM potassium ferricyanide (III) anhydrous in phosphate buffer 0.2 M at pH 7.4 ± 0.2 (dissolve 4.415 g/L NaH2PO4, 22.5075 g/L Na2HPO4•Heptahydrate pH should be 7.4, if not adjust with 1.0 N NaOH or 1.0 N HCl) and 0.5 g/L NaN3. Filter if necessary and store in a brown glass bottle or in dark plastic bottle at 4 °C.
Mix 0.2 ml of serum or cell lysates, through twice cycle of frozen and thawed, with 0.8 ml of substrate buffer and incubate for 1 hr at 37 °C, then add 1 ml blocking reagent, mix and incubate for 10 min in the dark. In last, dispense 1 ml color reagent, mix again and incubate for 10 min. At the finish read Abs at 510 nm.
The inhibition of this activity was also measured after adding 0.05 ml of 1 M sodium L-tartrate to a sample
before its incubation for 1 h at 37 °C.
Acid phosphatase activity units were expressed as nM of phenol produced per minute per 106 cells
Reply With Quote

acid , estimation , phosphatase

Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
USA - custom petides? avinash Proteomics Forum 6 06-09-2010 04:10 AM
10 mM HCl for IGF-I low.d Chemistry Forum 19 01-22-2010 12:33 AM
Acid phosphatase Sakkthi subramaniyam Protocols and Methods Forum 1 05-15-2009 04:49 PM
Deposizione rame su tubi in alluminio Max83 Forum Chimica 18 01-12-2008 05:48 PM
Testing Nitric Acid Etchant Bob Davis Chemistry Forum 0 12-26-2003 09:36 AM

All times are GMT. The time now is 11:26 PM.

Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2015, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.12863 seconds with 16 queries