CHEMISTRY REAGENT KITS
                          ------->MICRO PROTEIN KIT (Turbidometry)
                                                               For the determination of Proteins in urine and CSF

                                                                                      DOWNLOAD PDF

CLINICAL SIGNIFICANCE
Proteins are involved in the maintenance of the normal distribution of water between Wood and the tissues and consist mainly of the albumin and globulin fractions. The measurement of tow levels of urinary proteins is important in the detection of renal diseases. Proteinurea occurs in increased glomerular permeability and defective tubular reabsorption. Albuminuria is recognised as an early indicator or reversible renal damage in diabetics. The measurement of CSF proteins is used for the detection of increased permeability of the Dtoodybrain barrier in various diseases.

PRINCIPLE
In the presence of sufphosalicySc acid and sodium sulphate, Proteins yield a uniform turbidity which absorbs maximum at 546 nM or GREEN filter and is directly proportional to the concentration of Proteins.

REAGENTS SUPPUED
Cat No. ES 377 2x25 mL
1. CSF Protein Reagent 2x25 mL 5-
    Sulphosalicylic Acid 120 mmol/L Sodium
Sulphate 500 mmolt
2. Standard 5mL (Albumin Fraction V)
Contains stabilisers and sodium azide as
preservative.

Preparation of Working Reagent
Reagents are ready to use. Protect from bright light
The High Sensitivity Assay dilute the Standard(S) 1 +4 with normal saline. Prepare fresh each time.

STORAGE & STABILITY
Contents are stable at 2-8 C till the expiry
mentioned on the labels.

SAMPLE
Non-traumatic CSF or Urine. Sample must be free from visible turbidity. No prior patient preparation is required. If Urine to be assayed use fresh urine only. (All samples should be handled as potential infective agents as no laboratory methods make conclusive findings for its safety. Therefore, adequate protective laboratory measures should be taken while handling such materials).

SYSTEM PARAMETERS

Reaction End Point
Wavelength 530 nM Or 546 nM
(Gr»«i FUefl"
ZeroSetting Reagent Blank
Incubation Temperatue R.T.
Incubation Time 10 min.
Reagent Volume 500 µL  1000 µL
Sample Volume 50 µL  100 µL
aarxfardO>rK»ntrat)on OOmg'dL
Reaction Slope Increasing
Linearity 200 mg/dL
Units     mg/dL

PROCEDURE
Primary Wavelength / Filter          546 nM (Green Filter)
Secondary Wavelength / Filter      ' 0'
Temperature                                 R.T.
Light Path                                     1 cm.

MANUAL METHOD
Pipette into clean dry test tubes labeled as Blank (B), Standard (S), and Test (T).
    Label 3 Test tubes as B
(ml)
S
(ml)
T
(ml)
    Micro Protein Reagent - 1 3.0 3.0 3.0
    Distilled Water 0.2 - -
    Standard Reagent-2 - 0.2 -
    Sample - - 0.2

Mix well and incubate at R.T. for 10 minutes. Measure the absorbance of the Standard (Abs.S) and Test Sample (Abs.T) against the Blank, within 30 minutes.

MICRO METHOD FOR ANALYZERMICRO
Pipette into clean dry test tubes labeled as Blank (B), Standard (S), and Test (T)
    Label 3 Test tubes as B
(ml)
S
(ml)
T
(ml)
    Micro Protein Reagent - 1 1.0 1.0 1.0
    Distilled Water 0.1 - -
    Standard Reagent - 2 - 0.1 -
    Sample - - 0.1
Mix well and incubate at R.T. for 10 minutes. Measure the absorbance of the Standard (Abs.S) and Test Sample (Abs.T) against the Blank, within 30 minutes.Avoid using Dichromatic mode.

NORMAL REFERENCE VALUES
Urine   :   20-140 mg/24 hours
CSF     :   < 40 mg/dL

It is recommended that each laboratory establish Its own normal range representing its patient population.

CALCULATIONS
                                            Abs.T
Micro Proteins in mg/dL=   ---------- x 100 mg/dL
                                            Abs.S

LINEARITY
TTre Tea is linear ipto 200 mg/dL if values exceed this limit, dilute the sample with distilled water and repeat the assay. Multiply the value using eg. 2 for1:1dilutionfaclor.

WARNING
This reagent system is tor in vitro use only. This reagent system contains preservatives and components that have not established lor salety if contacted on broken skin or eye or taken oraly. In case ot such incidents wash off with plenty of water, or consult a physician.

QUALITY CONTROL
To ensure adequate quaity control, each kit should be tested against a standard control sera. I! should be realised that the use of quality control material checks both instrument and reagent function together. Fac^ which rr^aflectlne performance ol this test include proper instrument (unction, temperature control, cleanliness of glasswares and accuracy of pippettJng.
It is appropriate to establish each laboratory's accuracy constant and interpret values accordingly. Similarly, laboratory findings should be established by clinical manifestations.

BIBLIOGRAPHY
1.KING EJ. AND HASLEWOOD, GAD (1936), Lancet, 2. 1153.
2.KINGSBURG, F.P.. CLARK C.P.. WILLIAMS
3.AND POST A.I. (1926) J. Lab. Clin. Med. 11981.