ACPP ELISA Kit

Catalog No. ABIN996895

Product Details ACPP ELISA Kit

Target: ACPP (Acid Phosphatase, Prostate (ACPP))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0-60 ng/mL

Minimum Detection Limit: 0 ng/mL

Application: ELISA

Purpose: For the quantitative measurement of human prostatic acid phosphatase (PAP) in human serum and plasma.

Sample Type: Serum

Analytical Method: Quantitative

Specificity: 98.7%

Sensitivity: 0.2 ng/mL

Application Details

Comment:

Limitations of procedure:
1. The PAP values should be used as an adjunct - other data available - the physician.
2. Sample with PAP level above 60 ng/mL should be diluted - obtain an accurate value.

Sample Volume: 50 μL

Assay Time: 1 - 2 h

Plate: Pre-coated

Protocol: 1. It is important to wash microwells thoroughly and remove the last droplets of water to achieve the best results.
2. Pipette all reagents and samples into the bottom of the wells.
3. Absorbance is a function of time and temperature of incubations. It is recommended to have all reagents and sample caps removed and all needed wells assigned and secured in holder. It will ensure the equal elapsed time for each pipetting without interruption.

Reagent Preparation:

1. Bring all reagents and specimens to room temperature (20-25 °C) and gently swirl to obtain thorough mixing.
   2. Have all reagents and samples ready before the start of the assay. Once the test is begun, it must be performed without interruption to get the most reliable and consistent results.
   3. Prepare 1 x washing buffer.
   4. Prepare washing buffer by adding distilled or deionized water to 20x wash concentrate to a final volume of 1 liter.

Sample Preparation:

1. Collect blood specimens and separate the serum.
   2. Specimen may be refrigerated at 2-8 °C for up to seven days or frozen for up to six months. Avoid repetitive freezing and thawing of serum samples.

Assay Procedure:

1. Place the desired number of coated strips into the holder.
   2. Dispense 50 μL of standards and specimens into the appropriate wells.
   3. Dispense 50 μL of Sample diluent to each well and incubate for 30 minutes at room temperature.
   4. Remove incubation solution and wash three times with wash buffer.
   5. Dispense 100 μL of enzyme conjugate, to each well and incubate for 30 minutes at room temperature.
   6. Remove incubation solution and wash three times with wash buffer.
   7. Dispense 100 μL of TMB Chromogenic Substrate to each well and incubate for 15 minutes at room temperature.
   8. Add 100 μL of Stop solution to stop reaction.
   9. Read O.D. at 450 nm with a microwell reader.

Calculation of Results:

Construct a standard curve by plotting O.D. 450 nm on the y-axis against the concentration of PAP ng/mL on the x-axis using linear graph paper or log-log graph paper. Using the O.D. value of each specimen, determine the concentration of PAP from the standard curve. An example of typical results: Standard Set PAP (ng/mL) O.D. 450 nm O.D. 450 nm Mean SD CV % Standard 1 0 0.046 0.041 0.044 0.004
8.128 Standard 2
1.5 0.108 0.099 0.104 0.006
6.149 Standard 3 5 0.302 0.306 0.304 0.003 0.930 Standard 4 15 0.773 0.778 0.776 0.004 0.456 Standard 5 30
1.501
1.464
1.483 0.026
1.765 Standard 6 60
3.003
3.059
3.031 0.040
1.306 Control 1 -
1.740 0.116 0.113 0.115 0.002
1.853 Control 2 - 27.591
1.232
1.229
1.231 0.002 0.172 0 5 10 15 20 25 30 35 40 45 50 55 60 PAP ng/mL
Accuracy (Recovery) Recovery studies were performed by mixing an aliquot of pooled serum and PAP standard. The PAP values were measured and percentage of recovery were determined. Initial values ng/mL Conc. Spiked ng/mL Expected values ng/mL Observed values ng/mL Recovery % A: 4 5
4.5
4.5 100 4 15
9.5
1.0 105 4 30 17.0 16.8 99 B:
1.8 5
7.9
8.3 105
1.8 15
12.9 1
3.0 101
1.8 30 20.4 20.5 99 C: 28 3 15.5 15.0 97 28 5 16.5 17.0 103 28 15 2
1.5 2
3.5 109 Parallelism ce Dilution Calculated (ng/mL) Observed (ng/mL) Recovery % 4 in 4 86.0 3 in 4 64.5 69.7 108 2 in 4 4
3.0 45.2 105 1 in 4 2
1.5 2
1.0 98 Precision (Reproducibility) Inter-assay (n=12) and intra-assay (n=12), coefficient of variation, were evaluated at three different pooled serum samples: Inter-assay Intra-assay Pool A Pool Pool B C PoolA Pool B Pool C N 12 12 12 12 12 12 Mean (ng/mL) S.D. (ng/mL)
3.73
9.91 18.7
5.62
11.21 2
2.75 0.20 0.98 0.96 0.23 0.62 0.94 C.V. %
5.28
9.92
5.15
4.15
5.57
4.14

1. It is recommended that each laboratory determine its own normal value and abnormal range.

2. The results of a clinical study using ELISA PAP were summarized: Serum samples from 95 normal subjects were assayed. In this population, 99 % of the values was less than 3 ng/mL. a C Samples from 69 patients with benign prostatic hypertrophy (BPH) were assayed and found 94 % less than 3 ng/mL. b. Samples from 43 patients with prostatic carcinoma (PC) were assayed and found 81 % higher than 3 ng/mL. c

Restrictions: For Research Use only

Handling

Storage: 4 °C

Expiry Date: 12-14 months

Target Details for ACPP

Target: ACPP (Acid Phosphatase, Prostate (ACPP))

Alternative Name: Prostatic Acid Phosphatase (PAP) (ACPP Products)

Synonyms: 5'-NT ELISA Kit, ACP-3 ELISA Kit, ACP3 ELISA Kit, PAP ELISA Kit, AMAP2 ELISA Kit, CENTB3 ELISA Kit, DDEF2 ELISA Kit, PAG3 ELISA Kit, Pap-alpha ELISA Kit, SHAG1 ELISA Kit, A030005E02Rik ELISA Kit, FRAP ELISA Kit, Lap ELISA Kit, Ppal ELISA Kit, Acpp11 ELISA Kit, RNACPP11 ELISA Kit, pap ELISA Kit, ACPP ELISA Kit, acp-3 ELISA Kit, acp3 ELISA Kit, acpt ELISA Kit, acid phosphatase, prostate ELISA Kit, ArfGAP with SH3 domain, ankyrin repeat and PH domain 2 ELISA Kit, prostatic acid phosphatase ELISA Kit, prostatic acid phosphatase, putative ELISA Kit, acid phosphatase, prostate S homeolog ELISA Kit, ACPP ELISA Kit, ASAP2 ELISA Kit, Acpp ELISA Kit, CpipJ_CPIJ004002 ELISA Kit, Smp_016640 ELISA Kit, acpp.S ELISA Kit

Background: Prostatic acid phosphatase (PAP) enzyme activity was first measured in the urine of men and was found to be localized in organs of the male genital tract. Gutman and co-workers suggested that PAP may be a significant tumor marker in those patients with prostate cancer because serum PAP concentrations were found to be elevated in many men with primary prostatic carcinoma and metastatic lesions of prostate. In 1938, Gutman and Gutman reported elevated serum acid phosphatase activity in prostatic cancer patients, especially those with bone metastasis. Subsequent studies confirmed that this increased enzyme activity was of prostatic origin, also, the properties of this prostatic enzyme differed from those of acid phosphatase in normal serum. For many years serum acid phosphatase has been measured by spectrophotometric assays based on enzyme activity. These colorimetric methods utilize various substrates, some in conjunction with differential inhibitors of prostatic acid phosphatase. Generally, these assays lack sensitivity or specificity, also, the stability of serum enzymatic activity is time, temperature, and pH dependent. ELISA has been developed to provide a method of high sensitivity and specificity.

Pathways: Synaptic Membrane, Ribonucleoside Biosynthetic Process