Patent US4748115A successfully refreshed from Google Patents.
US4748115A
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US
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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
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 16:26
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 |