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US4748115A Expired / Lapsed US ⏱️ 0.0 years remaining

Substrate formulation in 2-amino-2-methyl-1-propanol buffer for alkaline phosphatase assays

Assignee: Abbott Laboratories
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Filing: 1986-01-08
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Granted: 1988-05-31
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Expires: 2006-01-08
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Abstract

Abstract A stable substrate formulation for an alkaline phosphatase assay includes 5-bromo-4-chloro-3-indolyl phosphate, nitro blue tetrazolium and a 2-amino-2-methyl-1-propanol buffer. Addition of MgCl 2 to this formulation promotes stability.

Inventors

  • 👤 Jeffrey W. Steaffens

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-11-03 00:46
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/34; C12Q1/42; C12Q2326/00; C12Q2326/90; C12Q2326/92; C12Q2334/00; C12Q2334/50; C12Q2334/52; Y; Y10; Y10S; Y10S435/00; Y10S435/967; Y10S436/00; Y10S436/805

Claims Analysis

Total Claims: 12
Claim 1
Independent Claim 65 words
A substrate formulation for an alkaline phosphatase assay comprising: a phosphated indigo cogener; a tetrazolium salt at a concentration effective to produce a detectable signal upon reaction with said phosphated indigo cogener; and 2-amino-2-methyl-1-propanol at a concentration effective to buffer a reaction between said phosphated indigo cogener and said tetrazolium salt in aqueous solution within a range of pH which allows the reaction to proceed.
Claim 2
Dependent Claim 65 words
A substrate formulation for an alkaline phosphatase assay comprising: a phosphated indigo cogener; a tetrazolium salt at a concentration effective to produce a detectable signal upon reaction with said phosphated indigo cogener; and 2-amino-2-methyl-1-propanol at a concentration effective to buffer a reaction between said phosphated indigo cogener and said tetrazolium salt in aqueous solution within a range of pH which allows the reaction to proceed.
Claim 3
Dependent Claim 16 words
The substrate formulation as recited in claim 1 wherein said phosphated indigo cogener is 5bromo-4-chloro-3-indolyl phosphate.
Claim 4
Dependent Claim 24 words
The substrate formulation as recited in claim 2 wherein said 5-bromo-4-chloro-3-indolyl phosphate is at a concentration between about 1.0 mM and about 10 mM.
Claim 5
Dependent Claim 16 words
The substrate formulation as recited in claim 1 wherein said tetrazolium salt is nitro blue tetrazolium.
Claim 6
Dependent Claim 25 words
The substrate formulation as recited in claim 4 wherein said nitro blue tetrazolium is at a concentration between about 0.05 mM and about 0.5 mM.
Claim 7
Dependent Claim 22 words
The substrate formulation as recited in claim 1 wherein said 2-amino-2-methyl-1-propanol is at a concentration between about 0.1M and about 1.0 M.
Claim 8
Dependent Claim 51 words
A substrate formulation in aqueous solution for an alkaline phosphatase assay comprising: 5-bromo-4-chloro-3-indolyl phosphate at a concentration between about 1.0 mM and about 10 mM.; nitro blue tetrazolium at a concentration between about 0.05 mM and about 0.5 mM; and 2-amino-2-methyl-1-propanol at a concentration between about 0.M and about 1.0 M.
Claim 9
Dependent Claim 51 words
A substrate formulation in aqueous solution for an alkaline phosphatase assay comprising: 5-bromo-4-chloro-3-indolyl phosphate at a concentration between about 1.0 mM and about 10 mM.; nitro blue tetrazolium at a concentration between about 0.05 mM and about 0.5 mM; and 2-amino-2-methyl-1-propanol at a concentration between about 0.M and about 1.0 M.
Claim 10
Dependent Claim 19 words
The substrate formulation as recited in claim 7, further comprising MgCl 2 at a concentration of about 1.0 mM.
Claim 11
Dependent Claim 40 words
The substrate formulation as recited in claim 8 wherein: said 5-bromo-4-chloro-3-indolyl phosphate is at a concentration of 1.2 mM; said nitro blue tetrazolium is at a concentration of 0.17 mM; and said 2-amino-2-methyl-1-propanol is at a concentration of 100 mM.
Claim 12
Dependent Claim 17 words
The substrate formulation as recited in claim 9, further comprising sodium azide at a concentration of 0.02%.

References Cited (Backward Citations)

US4030995A Cited
US4563417A Cited

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1991-05-31 1991-11-30 1992-05-31 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1995-05-31 1995-11-30 1996-05-31 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 1999-05-31 1999-11-30 2000-05-31 $7700.00 Window Closed / Lapsed

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