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

Product Details

Citations

Related Products

Customer Reviews and Q&A

Target Background

Name: Mouse tartrate-resistant acid phosphatase 5b(TRACP-5b) ELISA Kit
Brand: CUSABIO
SKU: CSB-E08492m

Product Details

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

Uniprot No.

Q05117

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

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.
	Sample	Serum(n=4)
1:1	Average %	100
	Range %	96-104
1:2	Average %	98
	Range %	92-107
1:4	Average %	93
	Range %	89-97
1:8	Average %	95
	Range %	90-99

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
EDTA plasma (n=4)	91	87-98

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
2.5	2.029	2.007	2.018	1.850
1.25	1.655	1.564	1.610	1.442
0.625	1.173	1.129	1.151	0.983
0.312	0.734	0.782	0.758	0.590
0.156	0.491	0.456	0.474	0.306
0.078	0.247	0.227	0.237	0.069
0	0.168	0.167	0.168

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

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

ELISA Data Analysis

ELISA Standard Curve & Curve Expert software

Troubleshooting
and FAQs

ELISA kit FAQs

Storage

Store at 2-8°C. Please refer to protocol.

Lead Time

3-5 working days after you place the order, and it takes another 3-5 days for delivery via DHL or FedEx

Description

This Mouse ACP5 ELISA Kit was designed for the quantitative measurement of Mouse ACP5 protein in serum, plasma, tissue homogenates. It is a Sandwich ELISA kit, its detection range is 0.078 mIU/mL-5 mIU/mL and the sensitivity is 0.02 mIU/mL.

Customer Reviews and Q&A

■ Customer Reviews

There are currently no reviews for this product.

Submit a Review here

■ Q&A

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.

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.

Target Background

Function
(From Uniprot)

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.

Gene References into Functions

TRAP and nidogen-2 co-localized in adipose tissue cells in situ. PMID: 25450682
Uteroferrin is a cytokine secreted by uterine glands in response to progesterone that promotes fetal erythropoiesis at various stages of pregnancy. PMID: 25093463
Supraphysiologic oscillatory fluid shear induces upregulation of osteoclastic activity, measured by tartrate-resistant acid phosphatase activity and formation of mineral resorption pits. PMID: 24275439
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
Serum TRAP levels were lower in mice receiving eggshell calcium citrate malate than those receiving eggshells calcium malate. PMID: 23603074
tartrate resistant acid phosphatase PMID: 23444125
TRAP is not essential for the formation of the epiphyseal vascular network. PMID: 22579636
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
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
USF1 and USF2 play a functional role in RANKL-dependent TRAP expression during osteoclast differentiation. PMID: 12663664
TRACP expression is a target for regulation by the macrophage/osteoclast transcription factor PU.1 and the osteoclast commitment factor MiTF PMID: 14584903
YY1 plays a functional role in RANKL-mediated TRAP gene expression during osteoclast differentiation. PMID: 15563837
distinct tartrate-resistant acid phosphatase promoter regions have distinct roles in gene regulation in transgenic mice PMID: 15590658
MMP-2 and MMP-9 expression by TRAP positive mononuclear and multinucleated cells are involved in articular cartilage destruction in CIA. PMID: 15862156
TRAP release is due to secretion, rather than cell death PMID: 16475168
TRACP is important for polarizing responses in naive T cells to antigen-presented dendritic cells. PMID: 16939395
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
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
prolonged hypoxia induces the formation of TRAP-positive osteoclast-like cells, suggesting the occurrence of an autocrine mechanism for osteoclastogenesis PMID: 18056037
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
central role for ACP5 in removal of the Man-6-P recognition marker from lysosomal proteins, which are then dephosphorylated PMID: 18940929
in the absence of RANKL and osteoclast differentiation, STAT6 binds the TRAP promoter after IL-4 treatment and directly enhances TRAP expression PMID: 19801646

Hide All

Subcellular Location

Lysosome.

Tissue Specificity

Characteristic constituent of osteoclasts.

Database Links

KEGG: mmu:11433

STRING: 10090.ENSMUSP00000065425

UniGene: Mm.46354

Code	CSB-E08492m
Size	96T,5×96T,10×96T
Price	Request a Quote or Start an on-line Chat
Trial Size	24T ELISA Kit Trial Size (Only USD$150/ kit) * The sample kit cost can be deducted from your subsequent orders of 96T full size kits of the same analyte at 1/5 per kit, until depleted in 6 months. Apply now

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.