Rat Phosphorylated tau, P-Tau ELISA Kit

Catalog No.: BZEK2044-48

Size: 48T

Range: 10 ng/L - 160 ng/L

Sensitivity: 1 ng/L

Storage and Expiration: Store at 2-8℃ for 6 months. 

Application: For quantitative detection of P-Tau in rat serum, plasma, tissue homogenate or cell culture supernatant.

Introduction

Tau, also known as MAPT (microtubule-associated protein tau), MAPTL, MTBT1 or TAU, is a 758 amino acid protein that localizes to the cytoplasm, as well as to the cytoskeleton and the cell membrane, and contains four Tau/MAP repeats. Expressed in neuronal tissue and existing as multiple alternatively spliced isoforms, Tau functions to promote microtubule assembly and stability and is thought to be involved in the maintenance of neuronal polarity. Tau may also link microtubules with neural plasma membrane components and, in addition to its role in microtubule stability, is also necessary for cytoskeletal plasticity. Tau is highly subject to a variety of post-translational modifications, including phosphorylation on serine and threonine residues, polyubiquitination (and subsequent proteasomal degradation) and glycation of specific Tau isoforms.

Principle of the Assay  

This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti- P-Tau antibody was pre-coated onto 48-well plates. And the HRP conjugated anti- P-Tau antibody was used as detection antibodies. The standards, test samples and HRP conjugated detection antibody were added to the wells subsequently, mixed and incubated, then, unbound conjugates were washed away with wash buffer. TMB substrates (A & B) were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the P-Tau amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of P-Tau can be calculated.

Kit components

1. One 48-well plate pre-coated with anti- rat P-Tau antibody

2. Standard: 0.5ml (320 ng/L)

3. Standard diluent buffer: 1.5 ml

4. Wash buffer (20×): 20 ml. Dilution: 1:20

5. Sample diluent buffer: 3 ml

6. HRP conjugated anti- rat P-Tau antibody (RTU): 3ml

7. Stop solution: 3 ml

8. TMB substrate A:  3ml

9. TMB substrate B:  3ml

10. Plate sealer:   2

11. Hermetic bag:  1

Material Required But Not Provided

1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Absorbent filter papers
8. Plastic or glass container with volume of above 1L