LEGEND:
▀ Substrate ▀ Buffer ▀ Indicator ▀ Coenzyme ▀ Activator ▀ Cofactor
RGT 1 RGT 2
METH
COMPONENT FUNCTION COMPONENT FUNCTION
Imidazole / Good’s Buffer ● (pH 6.8 – 6.9) Imidazole/Good’s buffer ● (pH 6.8 – 6.9)
(120mmol/L) ● Buffer (90mmol/L) ● Buffer
● Substrate for the 2nd reaction, ● Source of phosphate;
Glucose ADP
oxidation rgnt. for intermed. Rxn ● Substrate of the 1st rxn
(25mmol/L) (10mmol/L)
● Phosphate acceptor
● Reactive the enzyme bec a ● Inhibit interference of
prolonged standing your enzyme Adenylate kinase
N-acetylcysteine AMP
will oxidize – rendering it
(25mmol/L) (28mmol/L)
inactive to reverse inhibition of
sulfhydryl group
● Cofactor/ activator ● Indicator enzyme
● Alter the spatial configuration, it ● Source of hydrogen
may change the shape of the Glucose-6-phosphate
CK-MB FS Magnesium acetate
active site of the enzyme, to fit dehydrogenase
Oliver Rosalki (12.5mmol/L)
the substrate (≥ 15kU/L)
● Alters spatial configuration of
enzyme for stronger binding
● Preservative ● Indicator enzyme
EDTA-Na2 Diadenosine pentaphosphate
● Inhibitor; too much activator can ● Inhibit interference of
(2mmol/L) (50μmol/L)
inhibit the enzymes Adenylate kinase
● Coenzyme ● Substrate
NADP ● Hydrogen acceptor Creatine phosphate ● Source of phosphate on the first
(2.5mmol/L) ● 3rd reaction; come from vitamin (150mmol/L) rxn
B (niacin)
● Enzyme responsible for
Hexokinase
intermed. Rxn. Stabilizers
(≥ 5kU/L)
● The enzyme that catalyzes the
LEGEND:
▀ Substrate ▀ Buffer ▀ Indicator ▀ Coenzyme ▀ Activator ▀ Cofactor
2nd rxn
● Source: the goat
Monoclonal antibodies against
● Inhibit CK-M since CK-B is the
human CK-M
one determined to eliminate the
(2500U/L)
isoenzyme CK - MM
N-methyl-D-glucamine ● (pH 8.8- 9.8) NAD+ ● Coenzyme
LDH FS (420mmol) ● Buffer (50mmol/L) ● Hydrogen acceptor
Amador et al,
Wacker, Ulmer, ● Substrate
L-lactate
Valle ● Source of hydrogen; hydrogen
(65mmol/L)
donor
TRIS buffer ● (pH 7.65) 2-oxoglutarate ● Amino acid acceptor
(110mmol/L) ● provides the pH (65mmol/L) ● Alpha keto acid
L-aspartate ● Substrate of transamination rxn. NADH ● Reducing substance
(329mmol/L) ● Source of amino group (1mmol/L) ● Coenzyme of the indication rxn
● Indicator enzyme ● Supplementary reagent
MDH
Pyridoxal-5-phosphate FS ● Coenzyme for all the
(≥ 800U/L)
transamination reaction
ASAT
UV Method by ● Isoenzyme ● (pH9.6;)
Karmen; ● Catalyst
Henry’s Mod ● Accelerator
LDH Good’s buffer
● Prevent interference of
(1200U/L) (100mmol/L)
pyruvate (which is normally
present in your serum sample)
● Indicator enzyme
● Coenzyme
Pyridoxal-5-phosphate ● Accepts amino acid and transfer
(13mmol/L) it to COO- in alpha keto acid
● Active form of vitamin B6
ALAT TRIS buffer ● (pH 7.15) 2-oxoglutarate ● Amino acid acceptor
LEGEND:
▀ Substrate ▀ Buffer ▀ Indicator ▀ Coenzyme ▀ Activator ▀ Cofactor
UV Method by (140mmol/L) ● Provides the pH (85mmol/L)
Karmen;
Henry’s Mod L-alanine ● Substrate of transamination rxn. NADH ● Reducing substance
(700mmol/L) (1mmol/L)
LDH ● Isoenzyme ● Supplementary reagent
Pyridoxal-5-phosphate FS
(≥ 2300U/L) ● Indicator enzyme
Good’s buffer ● (pH9.6)
(100mmol/L)
Pyridoxal-5-phosphate ● Coenzyme
(13mmol/L)
Good’s buffer ● (pH 7.15) Good’s buffer ● (pH7.15;
( 0.1mol/L) (0.1mol/L)
● Cofactor ● 4,6-ethylidene-(G7)-p-nitrophe
nyl-(G1)-α-D-maltoheptaoside
(EPS-G7) OR ETHYLIDINE
NaCl EPS-G7
PROTECTED SUBSTRATE
(62.5mmol/L) (8.5mmol/L)
● Substrate
● Prevent interference of residual
AMS activity
α-AMYLASE
● Cofactor
Complete Color
● Activator
Fluid Stable
● Sources of chloride which serves
Mg2Cl
as allosteric activator (changes
(12.5mmol/L)
spatial configuration of the
enzyme for better substrate
binding)
● Indicator enzyme
● Auxiliary enzymes
α-glucosidase
● Redox enzymes
(≥ 2kU/L)
● Acts as a conjunction with the
protein enzyme
LEGEND:
▀ Substrate ▀ Buffer ▀ Indicator ▀ Coenzyme ▀ Activator ▀ Cofactor
● (pH 8.0) ● (pH.4.0)
Good’s buffer Tartrate buffer
● Alk – optimum pH ● Better dissolution of subs.
(50mol/L) (7.5mmol/L)
micro emulsion
LIPASE Taurodesoxycholate ● Bile salt ● Bile salt
Direct Color (4.3mmol/L); ● Accelerator Taurodesoxycholate ● Accelerator
Fluid Stable Desoxycholate ● Emulsifier (17.2mmol/L) ● Emulsifier
Enzymatic (8.0mmol/L)
Colorimetric
Method Calcium chloride ● Activator Color substrate ● (6-methylresorufin ester)
(15mmol/L) (0.65mmol/L) ● Substrate
Colipase ● Coenzyme ●
(2.2mg/L)
2-amino-2-methyl-1 propanol ● (pH10.4) P-nitrophenylphosphate ● Substrate
(1.1mol/L) ● Phosphate acceptor (80mmol/L)
Magnesium acetate ● Activator (stability)
(2mmol/L) ● Source of magnesium
● Activator
Zinc sulfate ● Has high affinity for binding
(0.5mmol/L) (10x) to magnesium if in excess
● Source of zinc
Alkaline
Phosphatase ● Metal ion buffer
Fluid Stable ● Chelating agent
● Binds with zinc & magnesium
(inhibits zinc if in excess)
● Metal ions which buffers the
HEDTA
system to maintain optimal
(2.5mmol/L)
concentration of your zinc and
magnesium; the metal ion buffer
also chelates their potential
inhibitory ions which may be
present in the sample
LEGEND:
▀ Substrate ▀ Buffer ▀ Indicator ▀ Coenzyme ▀ Activator ▀ Cofactor
BUFFERS AND pH
METHODOLOGY BUFFER pH
CK-MB FS
Imidazole / Good’s Buffer 6.8 - 6.9 (nice)
Oliver Rosalki
LDH FS
N-methyl-D-glucamine 8.8 - 8.9
Amador et al, Wacker, Ulmer, Valle
ASAT R1 TRIS buffer 7.65
UV Method by Karmen; Henry’s
Mod R2 Good’s buffer 9.6
ALAT R1 TRIS buffer 7.15
UV Method by Karmen; Henry’s
Mod R2 Good’s buffer 9.6
α-AMYLASE
Good’s buffer 7.15
Complete Color Fluid Stable
LIPASE R1 Good’s buffer 8.0
Direct Color Fluid Stable
Enzymatic Colorimetric Method R2 Tartrate buffer 4.0
Alkaline Phosphatase
HEDTA
Fluid Stable
