| Description |
The Mouse tartrate-resistant acid phosphatase 5b (TRACP-5b) ELISA Kit is designed to measure TRACP-5 in biological samples, including serum, plasma, and tissue homogenates, in a quantitative fashion. This kit has been verified by multiple tests with advantages of high sensitivity, strong specificity, precision less than 10%, high recovery, and lot-to-lot consistency. Based on the Sandwich-ELISA technique in combination with the enzyme-substrate chromogenic reaction as well as colorimetric measurement, the levels of TRACP-5b in the sample can be calculated. TRACP-5b (or called ACP5), highly and specifically expressed in osteoclasts, is an isoform of metalloenzyme TRAP. It is a dimer derived from TRACP-5a by proteolytic cleavage of a repressive loop domain and is the enzymatically more active form. TRACP-5b is a sensitive and specific marker of bone resorption and bone remodeling in patients with rheumatoid arthritis (RA), chronic kidney disease (CKD), and cancer with bone metastasis. A... Read more |
| Target Name |
acid phosphatase 5, tartrate resistant |
| Alternative Names |
Acp5 ELISA Kit; T5ap ELISA Kit; Trap ELISA Kit; Tartrate-resistant acid phosphatase type 5 ELISA Kit; TR-AP ELISA Kit; EC 3.1.3.2 ELISA Kit; Tartrate-resistant acid ATPase ELISA Kit; TrATPase ELISA Kit; Type 5 acid phosphatase ELISA Kit |
| Abbreviation |
ACP5 |
| Species |
Mus musculus (Mouse) |
| Sample Types |
serum, plasma, tissue homogenates |
| Detection Range |
0.078 mIU/mL-5 mIU/mL |
| Sensitivity |
0.02 mIU/mL |
| Assay Time |
1-5h |
| Sample Volume |
50-100ul |
| Detection Wavelength |
450 nm |
| Research Area |
Signal Transduction |
| Assay Principle |
quantitative |
| Measurement |
Sandwich |
| Precision |
| Intra-assay Precision (Precision within an assay): CV% |
| Three samples of known concentration were tested twenty times on one plate to assess. |
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| Inter-assay Precision (Precision between assays): CV% |
| Three samples of known concentration were tested in twenty assays to assess. |
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| Linearity |
| To assess the linearity of the assay, samples were spiked with high concentrations of mouse TRACP-5b in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay. |
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Sample |
Serum(n=4) |
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| 1:1 |
Average % |
100 |
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| Range % |
96-104 |
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| 1:2 |
Average % |
98 |
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| Range % |
92-107 |
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| 1:4 |
Average % |
93 |
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| Range % |
89-97 |
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| 1:8 |
Average % |
95 |
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| Range % |
90-99 |
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| Recovery |
| The recovery of mouse TRACP-5b spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section. |
| Sample Type |
Average % Recovery |
Range |
|
| Serum (n=5) |
97 |
92-102 |
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| EDTA plasma (n=4) |
91 |
87-98 |
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| Typical Data |
| These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed. |
| mIU/ml |
OD1 |
OD2 |
Average |
Corrected |
|
| 5 |
2.412 |
2.464 |
2.438 |
2.270 |
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| 2.5 |
2.029 |
2.007 |
2.018 |
1.850 |
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| 1.25 |
1.655 |
1.564 |
1.610 |
1.442 |
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| 0.625 |
1.173 |
1.129 |
1.151 |
0.983 |
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| 0.312 |
0.734 |
0.782 |
0.758 |
0.590 |
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| 0.156 |
0.491 |
0.456 |
0.474 |
0.306 |
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| 0.078 |
0.247 |
0.227 |
0.237 |
0.069 |
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| 0 |
0.168 |
0.167 |
0.168 |
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| Materials provided |
- A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-mouse TRACP-5b antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
- Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
- One vial Biotin-labeled TRACP-5b antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
- One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
- One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
- One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
- One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
- One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
- One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
- One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
- Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
- An instruction manual
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| Materials not provided |
- A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
- An incubator can provide stable incubation conditions up to 37°C±5°C.
- Centrifuge
- Vortex
- Squirt bottle, manifold dispenser, or automated microplate washer
- Absorbent paper for blotting the microtiter plate
- 50-300ul multi-channel micropipette
- Pipette tips
- Single-channel micropipette with different ranges
- 100ml and 500ml graduated cylinders
- Deionized or distilled water
- Timer
- Test tubes for dilution
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| Storage |
Store at 2-8°C. Please refer to protocol. |
| Lead Time |
3-5 working days |
