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Alpha-2-Macroglobulin

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Alpha-2-Macroglobulin Albumin Modular P Application Note Guideline for Determination of Albumin in Serum/Plasma on Modular P General information Intended use The Application Note is intended for the quantitative determination of albumin in human sample material by turbidimetry on Modular P (1, 2). Measuring range Approximately 7.7-61 g/L depending on the specific lot of the calibrator. In case of post-concentration or -dilution the range can be expanded to 4-70 g/L. Reference interval 35-52 g/L (3). It is recommended to determine the reference interval for the local population. Instrument settings Instrument programming is performed according to “Instrument Settings” on page 3. Code No. Name Reagents Antibody Q 0328 DakoCytomation Polyclonal Rabbit Anti-Human Serum Albumin Reaction buffer S 2006 DakoCytomation Reaction Buffer 3 Diluent S 2005 DakoCytomation Dilution Buffer 1 Calibrator X 0908 DakoCytomation Human Serum Protein Calibrator Controls X 0939 DakoCytomation Human Serum Protein Low Control Samples Human serum, heparin-plasma or EDTA-plasma. Stable for 7 days at 2-8 ºC. Stable for 3 months at –20 ºC (if frozen only once). It is recommended to thaw frozen samples at 37 °C. Mix well before analysis. Calibrator Predilute the DakoCytomation Human Serum Protein Calibrator, code No. X 0908, 1:26.0 (e.g. 100 µL calibrator + 2500 µL diluent). The final dilution of standards is performed automatically by the instrument. Reaction buffer (R1) The reaction buffer is ready for use. On board stability is 28 days. Antibody (R3) Predilute the antibody 1:16.0 (e.g. 1200 µL antibody + 18000 µL diluent). If in rare cases the prediluted antibody appears slightly turbid, filtration through a 0.22 µm membrane filter is recommended. Stability of undiluted antibody: See expiry on the label. Stability of prediluted antibody: 28 days at 2-8 °C. On board stability: 28 days. Capacity: 1 mL of prediluted antibody is equivalent to approximately 18 cuvette readings of standards or samples. The dead volume of the reagent bottle should be considered when calculating the required amount of reagents. Calibration stability It is recommended to recalibrate every 28th day or when reagent lots change, a new antibody dilution is prepared, the antibody dilution is filtered, or quality control results fall outside the range as established by the individual laboratory. Trouble shooting If performance is unacceptable, try to recalibrate. Check reagents and procedure. If the problem persists, please contact instrument supplier or DakoCytomation Technical Service. (109498-002) 30 194.Q0328.01/UK/OS/11.01.05 p. 1/3 DakoCytomation Denmark A/S · Produktionsvej 42 · DK-2600 Glostrup · Denmark · Tel. +45 44 85 95 00 · Fax +45 44 85 95 95 · CVR No. 33 21 13 17 Albumin Modular P Performance Data Sensitivity An OD value of approximately 0.25 on Modular P corresponds to an albumin concentration around 60 g/L. Detection limit The detection limit is estimated to 0.1 g/L. Precision The precision was estimated by testing at 3 different albumin (ALB) levels by ANOVA analysis of 6 runs each with a new calibration and 6 determinations in each run. ALB Standard deviation (g/L) Mean Sera value Within Between Total CV (g/L) run run Total (%) n 1 38.8 0.47 1.69 1.76 4.5 36 2 41.3 0.64 0.73 0.97 2.3 36 3 95.6 1.59 2.90 3.31 3.5 32 Accuracy A recovery of albumin of 90-110% can be expected for DakoCytomation Human Serum Protein Low Control, code No. X 0939. Linearity The assay is linear in the range 7.7-61 g/L. Security range No antigen excess is found for albumin concentrations up to 298 g/L (the highest concentration tested). Interference No interference is found at concentrations up to 10 g/L of hemoglobin, 600 mg/L of bilirubin, and 25 g/L of triglyceride. All drugs described in reference 4 were investigated according to the recommendations in reference 4. No interference was observed. Method comparison Determination of albumin according to this Application Note was compared with other commercial turbidimetric assays. Data are available on request. References 1. Blirup-Jensen S. Protein Standardization III: Method optimization: Basic principles for quantitative determination of human serum proteins on automated instruments based on turbidimetry or nephelometry. Clin Chem Lab Med 2001;39:1098-1109. 2. Modular P manual(s). 3. Dati F, Schumann G, Thomas L, Aguzzi F, Baudner S, Bienvenu J, et al. Consensus of a group of professional societies and diagnostic companies on guidelines for interim reference ranges for 14 proteins in serum based on the standardization against the IFCC/BCR/CAP Reference Material (CRM 470). Eur J Clin Chem Clin Biochem 1996;34:517-20. 4. Sonntag O, Scholer A. Drug interference in clinical chemistry: recommendation of drugs and their concentrations to be used in drug interference studies. Ann Clin Biochem 2001;38:376-85. (109498-002) 30 194.Q0328.01/UK/OS/11.01.05 p. 2/3 DakoCytomation Denmark A/S · Produktionsvej 42 · DK-2600 Glostrup · Denmark · Tel. +45 44 85 95 00 · Fax +45 44 85 95 95 · CVR No. 33 21 13 17 Albumin Modular P Instrument Settings (Software Version 7675850-06-06) Analysis Assay/Time/Point 2 Point End 10 15 34 0 0 Wavelength (2nd/Primary) 800 340 Sample Volume Reagent Volume Normal 2 5 232 R1 250 0 5* 0 Timing Decrease 2 3 270 R2 0 0 5* 0 T2 Increase 4 5 230 R3 50 20 5* 0 T3 R4 0 0 5* 0 Diluent О Water Reagent Bottle Size О Diluent 952* 99 A 0* D 0* Abs. Limit 32000 Increase B 0* E 0* Prozone Limit 32000 0 0 0 0 Upper C 0* Cell Detergent Detergent 1 Twin Test Cancel Calibration Calibration Type Logit-Log 4P Auto Calibration Point 6 Time out Span 4 □ Blank 0 Weight 0 □ Span 0 Update Type None □ 2 Point 0 Isozyme Q Channel Cancel □ Full 0 SD Limit 50 Change over Duplicate Limit 20 % 100 Abs Module Cancel Sensitivity Limit -99999 999999 Lot Cancel S1 Abs. Limit -32000 32000 Bottle Cancel Range Application Code 5* Expected Value Unit g/L Male Report Name 100 Year -99999 999999 Albumin 100 Year -99999 999999 -99999 999999 Data Mode Active Female Technical Limit St1+0.001 Std. 6 100 Year -99999 999999 Repeat Limit Std. 1 Std. 6 100 Year -99999 999999 -99999 999999 □ Control Interval Time □ Qualitative Default (1) 0 -- Sex (2) 0 - О Male О Female (3) 0 +- (4) 0 ++ Range (5) 0 +++ О Range 1 О Range 2 О Range 3 (6) ++++ Others Standard (1) (2) (3) (4) (5) (6) Calibration Code* 501* 501* 501* 501* 501* 501* 4.5000 6.3243 9.4228 14.0000 18.0000 35.6087 Concentration*** x C(cal) x C(cal) x C(cal) x C(cal) x C(cal) x C(cal)1 Rack No. - Pos.* S001-3* S001-3* S001-3* S001-3* S001-3* S001-3* Sample volume 8 8 12 14 20 35 Diluted S. Volume 5 5 5 5 5 5 Diluent Volume 200 140 137 103 110 80 Diluent – Code No. S 2005 R1 – Code No. S 2006 R3 – Code No. Q 0328, diluted 1:16.0. [*] Defined by the customer. [**] The calculated relative concentration of Std.1 and Std.6. In order to get a warning for samples with concentration below the concentration of the lowest standard, the lower technical limit has to be 0.001 g/L higher than the lowest standard. [***] The prediluted calibrator (according to “Calibrator” on page 1), code No. X 0908. The concentration is calculated as the factor times the calibrator value for the specific lot (stated in g/L on the X 0908 Analytical Value Sheet). Number of decimals stated for Std.1 defines the number of decimals in printout. Two decimals are recommended. (109498-002) 30 194.Q0328.01/UK/OS/11.01.05 p. 3/3 DakoCytomation Denmark A/S · Produktionsvej 42 · DK-2600 Glostrup · Denmark · Tel. +45 44 85 95 00 · Fax +45 44 85 95 95 · CVR No. 33 21 13 17 .
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