Mouse tartrate-resistant acid phosphatase 5b(TRACP-5b) ELISA Kit

Datasheet
  • 中文名称:
    小鼠抗酒石酸酸性磷酸酶5b(TRACP-5b)ELISA Kit
  • 货号:
    CSB-E08492m
  • 规格:
    96T
  • 价格:
    3200
  • 其他:

产品详情

  • 产品描述:

    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. And TRAP-5b was proposed as a serum marker for bone metastases in various types of primary cancers.

  • 别名:
    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
  • 缩写:
    ACP5
  • Uniprot No.:
  • 种属:
    Mus musculus (Mouse)
  • 样本类型:
    serum, plasma, tissue homogenates
  • 检测范围:
    0.078 mIU/mL-5 mIU/mL
  • 灵敏度:
    0.02 mIU/mL
  • 反应时间:
    1-5h
  • 样本体积:
    50-100ul
  • 检测波长:
    450 nm
  • 研究领域:
    Signal Transduction
  • 测定原理:
    quantitative
  • 测定方法:
    Sandwich
  • 精密度:
    Intra-assay Precision (Precision within an assay): CV%<8%
    Three samples of known concentration were tested twenty times on one plate to assess.
    Inter-assay Precision (Precision between assays): CV%<10%
    Three samples of known concentration were tested in twenty assays to assess.
  • 线性度:
    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.
     SampleSerum(n=4)
    1:1Average %100
    Range %96-104
    1:2Average %98
    Range %92-107
    1:4Average %93
    Range %89-97
    1:8Average %95
    Range %90-99
  • 回收率:
    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 TypeAverage % RecoveryRange
    Serum (n=5) 9792-102
    EDTA plasma (n=4)9187-98
  • 标准曲线:
    These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
    mIU/mlOD1OD2AverageCorrected
    52.412 2.464 2.438 2.270
    2.52.029 2.007 2.018 1.850
    1.251.655 1.564 1.610 1.442
    0.6251.173 1.129 1.151 0.983
    0.3120.734 0.782 0.758 0.590
    0.1560.491 0.456 0.474 0.306
    0.0780.247 0.227 0.237 0.069
    00.168 0.167 0.168  
  • 本试剂盒所含材料:
    • 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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  • 本试剂盒不含材料:
    • 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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  • 货期:
    3-5 working days

引用文献

问答及客户评论

 常见问题解答
Q:

I’m searching for ELISA kits to detect TRAP in mouse plasma/serum and in culture supernatants of osteoclasts cultured on bovine bone slices.
I saw that Cusabio proposes a kit for the detection of TRAP with the reference CSB-E08492m.
Can you tell me if this kit is specific of TRAP5b produced by the osteoclasts (no detection of TRAP produced by other cells)?
Also, do you know if 100 μL of serum/plasma is necessary for the test or if a dilution of the samples is required?
Finally, as I would like to measure TRAP in culture supernatants, I was wondering if conditioned media (alpha-MEM containing inactivated FBS and penicillin/streptomycin) can be used as samples and if bovine TRAP coming from bone slices or FBS will be detected by the kit.
Thank you in advance for your consideration.

A:
Thanks for your inquiry.
Pls check our answer:
TRACP is a good marker of bone resorption and osteoclast activity.
TRACP especially in the presence of tracp-5a (mainly from macrophages) and tracp-5b (mainly from osteoclasts) existed in the blood.
The determination of tracp-5b can reflect osteoclast activity.
CSB-E08492m is specifically for TRACP-5b, properly used for the mouse matrix samples.
The common samples are serum and plasma. The sample adding volume is 100ul. There is no need to dilute.
As for the TRAP in culture supernatants, pls arrange the preliminary experiment. Pls kindly note detects the mouse TRACP-5b not TRAP.
Any question, pls feel free to contact me.

靶点详情

  • 功能:
    May play a role in the process of bone resorption. The osteoclastic trap acts on nucleotide tri- and diphosphates with higher affinity, compared with other substrates.
  • 基因功能参考文献:
    1. TRAP and nidogen-2 co-localized in adipose tissue cells in situ. PMID: 25450682
    2. Uteroferrin is a cytokine secreted by uterine glands in response to progesterone that promotes fetal erythropoiesis at various stages of pregnancy. PMID: 25093463
    3. Supraphysiologic oscillatory fluid shear induces upregulation of osteoclastic activity, measured by tartrate-resistant acid phosphatase activity and formation of mineral resorption pits. PMID: 24275439
    4. Pax6 binds endogenously to the proximal region of the tartrate acid phosphatase (TRAP) gene promoter and suppresses nuclear factor of activated T cells c1 (NFATc1)-induced TRAP gene expression. PMID: 23990468
    5. Serum TRAP levels were lower in mice receiving eggshell calcium citrate malate than those receiving eggshells calcium malate. PMID: 23603074
    6. tartrate resistant acid phosphatase PMID: 23444125
    7. TRAP is not essential for the formation of the epiphyseal vascular network. PMID: 22579636
    8. dephosphorylation of Man6P-containing lysosomal proteins requires the concerted action of Acp2 and Acp5 and is needed for hydrolysis and removal of degradation products PMID: 22158965
    9. Accumulation of vesicles in the cytoplasm of mutant osteoclasts indicates a novel function for tartrate-resistant acid phosphatase in modulating intracellular vesicular transport in osteoclasts PMID: 12243748
    10. USF1 and USF2 play a functional role in RANKL-dependent TRAP expression during osteoclast differentiation. PMID: 12663664
    11. TRACP expression is a target for regulation by the macrophage/osteoclast transcription factor PU.1 and the osteoclast commitment factor MiTF PMID: 14584903
    12. YY1 plays a functional role in RANKL-mediated TRAP gene expression during osteoclast differentiation. PMID: 15563837
    13. distinct tartrate-resistant acid phosphatase promoter regions have distinct roles in gene regulation in transgenic mice PMID: 15590658
    14. MMP-2 and MMP-9 expression by TRAP positive mononuclear and multinucleated cells are involved in articular cartilage destruction in CIA. PMID: 15862156
    15. TRAP release is due to secretion, rather than cell death PMID: 16475168
    16. TRACP is important for polarizing responses in naive T cells to antigen-presented dendritic cells. PMID: 16939395
    17. both synthesis as well as degradation of collagen are increased when TRAP is absent in mice at 8 weeks and 6 months of age, showing that TRAP has an important role in the metabolism of collagen PMID: 17551769
    18. transcriptional activity of the Tracp gene, in pre-osteoclastic cells, is negatively regulated by the binding of PARP-1 protein to a potential consensus sequence located in its promoter region PMID: 18021007
    19. prolonged hypoxia induces the formation of TRAP-positive osteoclast-like cells, suggesting the occurrence of an autocrine mechanism for osteoclastogenesis PMID: 18056037
    20. Over-expression of monomeric tartrate resistant acid phosphatase, but not the dimeric form in adipose tissue leads to early onset spontaneous hyperplastic obesity PMID: 18320034
    21. central role for ACP5 in removal of the Man-6-P recognition marker from lysosomal proteins, which are then dephosphorylated PMID: 18940929
    22. in the absence of RANKL and osteoclast differentiation, STAT6 binds the TRAP promoter after IL-4 treatment and directly enhances TRAP expression PMID: 19801646

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  • 亚细胞定位:
    Lysosome
  • 组织特异性:
    Characteristic constituent of osteoclasts.
  • 数据库链接: