CC2 LAB Flashcards
ASSAY ENZYME ACTIVITY:
Creatine Kinase
• TANZER-GILBARG (Forward/Direct)
• OLIVER-ROSALKI ( Reverse/Indirect)
ASSAY ENZYME ACTIVITY: CREATINE KINASE
- Forward/Direct
- pH : 9:0
- 340 nm
TANZER GIRLBARG
ASSAY ENZYME ACTIVITY: CK
- Reverse/Indirect
- pH : 6.8
- 340 nm
OLIVER-ROSALKI
1.1.1.27
LACTATE DEHYDROGENASE
1.1.1.49
GLUCOSE-6-PHOSPHATE DEHYDROGENASE
2.6.1.1
ASPARTATE AMINO-TRANSFERASE
2.6.1.2
ALANINE AMINO-TRANSFERASE
2.7.3.2
CREATINE KINASE
2.3.2.2
GAMMA-GLUTAMYL TRANSFERASE
3.1.3.1
ALKALINE PHOSPHATASE
3.1.3.2
ACID PHOSPHATASE
3.2.1.1
AMYLASE
3.1.1.8
ACETYL CHOLINESTERASE
3.1.1.3
LIPASE
3.1.3.5
5-NUCLEOTIDASE
What is the EC code no. of CK?
2.7.3.2
What is the EC code no. of LACTATE DEHYDROGENASE?
1.1.1.27
What is the EC code no. of GLUCOSE-6-PHOSPHATE DEHYDROGENASE?
1.1.1.49
What is the EC code no. of ASPARTATE AMINO-TRANSFERASE?
2.6.1.1
What is the EC code no. of ALANINE AMINO-TRANSFERASE?
2.6.1.2
What is the EC code no. of GAMMA-GLUTAMYL TRANSFERASE?
2.3.2.2
What is the EC code no. of ALKALINE PHOSPHATASE?
3.1.3.1
What is the EC code no. of ACID PHOSPHATASE?
3.1.3.2
What is the EC code no. of AMYLASE?
3.2.1.1
What is the EC code no. of ACETYL CHOLINESTERASE?
3.1.1.8
What is the EC code no. of 5-NUCLEOTIDASE?
3.1.3.5
What is the EC code no. of LIPASE?
3.1.1.3
A water-free cavity where the substrate interacts with particular charged amino acids residues.
ACTIVE SITE
Result when an enzyme is subject to posttranslational modifications.
ISOFORM
The non-protein molecule part of an enzyme
COFACTOR
Examples of Activators:
CHLORIDE & MAGNESIUM IONS
An organic cofactor such as NICOTINAMIDE ADENINE DINUCLEOTIDE
COENZYME
When a COENZYME is bound tightly to the ENZYME, the coenzyme is called what?
PROSTHETIC GROUP
PRINCIPLE: Catalyzes an oxidation-reduction reaction between two substrates
OXIDOREDUCTASES
PRINCIPLE: Catalyze the transfer of a group other than hydrogen from one substrate to another.
TRANSFERASES
PRINCIPLE: Catalyze hydrolysis of various bonds
HYDROLASES
PRINCIPLE: Catalyze the interconversion of geometric, optical, or positional isomers
ISOMERASES
PRINCIPLE: Catalyze the joining of 2 substrate molecules, coupled with breaking of the pyrophosphate bond
LIGASES
LIPASE SAMPLE
20 uL
LIPASE: reagent 1
1000 uL
LIPASE: Mode of reaction & slope of reaction
FIXED TIME; INCREASING
LIPASE: Delay time
120 secs
LIPASE
- What is the tissue source?
PANCREAS
MOST SPECIFIC and LATE MARKER for ACUTE PANCREATITIS
LIPASE
LIPASE 3 isoenzyme:
L1, L2, L3
Which among of the LIPASE ISOENZYME is the most chemically specific and sensitive?
L2
LIPASE ASSAY ENZYME ACTIVITY
- It measures the liberated fatty acids by titration after 24hr incubation
CHERRY CRANDAL
Use as a more pure form of triglyceride
TRIOLEIN
It catalyzes the breakdown of starch and glycogen into smaller sugars
AMYLASE
-It is the smallest enzyme
AMYLASE
- Requires CHLORIDE for activation
AMYLASE
Tissue sources of AMYLASE?
Acinar cells of the pancreas & salivary glands
2 ISOENZYME OF AMYLASE:
- S-TYPE
PTYALIN
It is the EARLIEST PANCREATIC MARKER
AMYLASE
2 ISOENZYME OF AMYLASE:
- S-TYPE (Ptyalin)
- P-TYPE (Amylopsin)
AMYLASE METHODOLOGIES:
- Coupling of several enzyme systems to monitor AMYLASE activity
CONTINUOUS MONITORING
AMYLASE METHODOLOGIES
- Measures the disappearance of starch substrate iodine.
AMYLOCLASTIC
2 ISOENZYME OF AMYLASE:
- P-type
AMYLOPSIN
AMYLASE METHODOLOGIES:
- Measures the appearance of the product
SACCHAROGENIC
AMYLASE:
- slope of reaction
INCREASING
AMYLASE METHODOLOGIES:
- Measures the increasing color from production of product coupled with a chromogenic dye.
CHROMOGENIC
AMYLASE REAGENT 1
1000 uL
Administration of ___________ and other/ opiates may cause false elevated serum
MORPHINE
Volume of sample for AMYLASE:
25 UL
AMYLASE SAMPLE:
25 uL
TISSUE SOURCES OF ACID PHOSPHATASE (ACP):
PROSTATE, LIVER, SPLEEN, RBC’s, PLATELETS & BONES
MAJOR ISOENZYMES OF ACP:
- PROSTATIC
- ERYTHROCETIC
- TARTRATE
ACP ISOENZYME:
- Resistant ACP
BONE
ACP ISOENZYME:
- Specific ACP; inhibited by TARTRATE
PROSTATIC
ACP ISOENZYME:
- Non-specific ACP; Inhibited by FORMALDEHYDE & CUPRIC SULFATE
ERYTHROCYTIC
ACP ASSAY ENZYME ACTIVITY:
- For quantitative endpoint reactions
THYMOLPHTHALEIN MONOPHOSPHATE
ACP ASSAY ENZYME ACTIVITY:
- Continuous monitoring methods
NAPHTHYL PHOSPHATE
ACP vol. of SAMPLE:
100 uL
ACP slope of reaction:
INCREASING
- Optimal pH for the reaction is 9-10
- requires MAGNESIUM as an ACTIVATOR
ALKALINE PHOSPHATASE
TISSUE SOURCES OF ALKALINE PHOSPHATASE (ALP):
INTESTINE, LIVER. BONE, PLACENTA, SPLEEN &B KIDNEY
ALP MAJOR ISOENZYMES:
INTESTINAL ALP, LIVER ALP, BONE ALP, PLACENTAL ALP
CARCINOPLACENTAL ALKALINE PHOSPHATASES:
REGAN & NAGAO
Often used in HEPATOBILIARY & BONE DISORDERS:
ALP
PHYSIOLOGIC ELEVATION OF ALP CAN BE SEEN IN?
PREGNANCY, CHILDREN (bone growth & development)
HIGHEST ELEVATION OF ALP is seen in _____________ Disease.
PAGETS DISEASE/OSTEITIS DEFORMANS
VOLUME OF SAMPLE FOR ALP:
20 uL
ALP SLOPE OF REACTION:
INCREASING
ASSAY ENZYME ACTIVITY FOR ALP:
BOWERS & MCCOMB
A continuous-monitoring technique that allows calculations of ALP activity based on the molar absorptivity of p-nitrophenol
BOWERS & MCCOMB
MOST LIVER SPECIFIC ENZYME
ALANINE AMINOTRANSFERASE
Other name for ALT?
SERUM GLUTAMIC-PYRUVIC TRANSAMINASE (SGPT)
Major source of ALT?
liver
DeRITIS ratio of ALT is >1.0
ACUTE HEPATITIS
COUPLED ENZYMATIC REACTION
- pH:
7.3-7.8
ASSAY ENZYME ACTIVITY FOR ALT:
COUPLED ENZYMATIC REACTION
REFERENCE RANGE FOR ALT:
6-37 U/L
SLOPE OF REACTION OF ALT:
DECREASING
Sample vol. of ALT:
100 uL
OTHER NAME FOR AST:
SERUM GLUMATIC-OXALOACETIC TRANSAMINASE (SGOT)
AST sources:
Cardiac tissue>liver>skeletal>muscle>kidney>pancreas>rbc
2 ISOENZYME OF AST:
- CYTOPLASMIC
- MITOCHONDRIAL
ASSAY ENZYME ACTIVITY for AST:
KARMEN METHOD
indicator ni KARMEN METHOD
MALATE DEHYDROGENASE
AST activity is stable in serum for ___ days at ___ temperatures
3-4 days ; refrigerated
slope of reaction of AST:
DECREASING
Sample vol. for AST TEST:
100 uLr
REFERENCE RANGE OF AST:
5-30 U/L
5 MAJOR ISOENZYMES OF LDH:
LD1 (HHHH), LD2 (HHHM), LD3 (HHMM), LD4 (HMMM), LD5 (MMMM)
Considered as LEAST SPECIFIC ENZYME
LACTATE DEHYDROGENASE
5 MAJOR ISOENZYMES OF LDH:
- Most anodal
LD1 (HHHH)
5 MAJOR ISOENZYMES OF LDH:
- Most Abundant
LD2 (HHHM)
LDH highest elevation is seen in?
PERNICIOUS/MEGALOSBLASTIC ANEMIA
ASSAY ENZYME FOR LDH:
- WACKER (forward/direct)
- WROBLEUSKI LADUE (reverse/indirect)
pH ni WACKER:
8.8
pH ni WROBLEUSKI LADUE:
7.2
REFERENCE RANGE OF LDH:
100-225 U/L
SLOPE OF REACTION OF LDH:
DECREASING
SAMPLE VOL. FOR LDH TEST:
10 u/L
3 MAJOR ISOENZYME: CK
Found in serum of HEALTHY INDIVIDUAL:
CK-MM (CK3)
Most commonly performed in the lab bcoz it takes 2-6 mins faster than tanzer
OLIVER-ROSALKI
OLIVER-ROSALKI pH:
6.8
TANZER-GILBARG pH:
9.0
ALP ISOENZYME FROM HEAT STABLE - HEAT LABILE
REGAN > NAGAO > PLACENTAL > INTESTINAL > LIVER > BONE
CARCINOPLACENTAL ALP
NAGAO
E.C. 3.1.1.6
PSEUDO-CHOLINESTERASE
Principle: HYDROLYZES BUTYRYL AND BENZYLCHOLINE
PSEUDO-CHOLINESTERASE
PSEUDOCHOLINESTERASE (PCHE)
- What is the SUBSTRATE in ELLMAN’S METHOD?
Acylthiocholine
TRUE OR FALLS
Because enzyme CATALYZE PHYSIOLOGIC REACTIONS, the ENZYME CONCENTRATION affects the rate of the CATALYZED REACTION.
TRUE
PROTEINS THAT CARRY NET MOLECULAR CHARGES
ENZYMES
Non protein entities that must bind to particular enzymes before a reaction occurs.
COFACTORS
COMMON ACTIVATORS:
(Ca2+, Fe2+,
Mg2+, Mn2+, Zn2+, and K+) and nonmetallic (Br- and Cl–).
E.C. 2.3.2.2
GAMMA-GLUTAMYL TRANSFERASE
CLINICAL SIGNIFICANCE:
Hepatobiliary disorders
- Patients receiving enzyme unducing drugs (warfarin, phenobarbital, phenytoin)
GAMMA-GLUTAMYL TRANSFERASE
SAMPLE for G-6-PD:
RED CELL HEMOLYSATE/SERUM
E.C.3.4.15.1
ANGIOTENSIN-CONVERTING ENZYME (AMS)
