TrkB 蛋白, Mouse (HEK293, His)

TrkB protein, also known as tropomyosin receptor kinase B, consists of a non-catalytic isoform. Unlike its catalytic isoform, the noncatalytic isoform of TrkB lacks intrinsic kinase activity. TrkB Protein, Mouse (HEK293, His) is the recombinant mouse-derived TrkB protein, expressed by HEK293 , with C-His labeled tag.

TrkB 蛋白，也称为原肌球蛋白受体激酶 B，由非催化亚型组成。与其催化亚型不同，TrkB 的非催化亚型缺乏内在激酶活性。TrkB 的催化亚型负责在其配体（例如脑源性神经营养因子 (BDNF)）激活后启动信号级联反应。然而，TrkB 的非催化异构体被认为可以调节催化异构体的活性并调节其信号通路。虽然 TrkB 非催化亚型的确切作用仍在研究中，但人们认为它在各种细胞过程中发挥调节作用，特别是在神经元发育和突触可塑性中。需要进一步的研究来充分阐明 TrkB 非催化亚型的功能意义及其对神经元功能的贡献。

1.Measured in a cell proliferation assay using SH-SY5Y human neuroblastoma cells. The ED₅₀ this effect is 0.3-0.6 μg/ml.
2.Measured by its binding ability in a functional ELISA. Immobilized recombinant Mouse/Human BDNF Protein (Native) at 10 μg/mL (100 μl/well) can bind biotinylated mouse TrkB-His with a linear range of 10-80 ng/mL.

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  Measured in a cell proliferation assay using SH-SY5Y human neuroblastoma cells. The ED₅₀ this effect is 0.5971 μg/mL, corresponding to a specific activity is 1674.76 units/mg.

Materials
Cells for assay: SH-SY5Y cells
Culture medium: Specialized culture medium
TrkB Protein, Mouse (HEK293, His) (HY-P72436)
Penicillin-Streptomycin (100×), Sterile (HY-K1006)
Cell Counting Kit-8 (HY-K0301)

Procedure
1. Seed cells at a density of 4000 cells per well in a 96-well plate, with a volume of 100 μL per well. (Do not seed cells in the outermost ring of the 96-well plate; instead, add an equal volume of PBS solution.)
2. After the cells have adhered, discard the culture medium in the wells and add 100 μL of specialized medium containing different concentrations of Mouse TrkB to the 96-well plate, resulting in final concentrations of 0, 0.001, 0.01, 0.05, 0.1, 0.25, 0.5, 1, 5, 20, 50, and 100 μg/mL in each well.
3. Incubate the cells at 37°C in a 5% CO₂ incubator for 48 hours.
4. After the cultivation period, add 10 μL of CCK8 solution to each well under light-protected conditions and incubate at 37°C in a 5% CO₂ incubator for 2 hours.
5. Measure the OD value at 450 nm using a microplate reader.
6. Using GraphPad Prism 9 software, plot a curve with OD values on the y-axis and protein concentration on the x-axis to calculate the ED₅₀ value.

Mouse

HEK293

C-His

P15209-1 (C32-H429)

18212  [NCBI]

Extracellular Domain

CPTSCKCSSARIWCTEPSPGIVAFPRLEPNSVDPENITEILIANQKRLEIINEDDVEAYVGLRNLTIVDSGLKFVAYKAFLKNSNLRHINFTRNKLTSLSRRHFRHLDLSDLILTGNPFTCSCDIMWLKTLQETKSSPDTQDLYCLNESSKNMPLANLQIPNCGLPSARLAAPNLTVEEGKSVTLSCSVGGDPLPTLYWDVGNLVSKHMNETSHTQGSLRITNISSDDSGKQISCVAENLVGEDQDSVNLTVHFAPTITFLESPTSDHHWCIPFTVRGNPKPALQWFYNGAILNESKYICTKIHVTNHTEYHGCLQLDNPTHMNNGDYTLMAKNEYGKDERQISAHFMGRPGVDYETNPNYPEVLYEDWTTPTDIGDTTNKSNEIPSTDVADQSNREH

Approximately 60-90 kDa due to the glycosylation.

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  Greater than 95% as determined by reducing SDS-PAGE.

无菌粉末

Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4 or 50 mM Tris-HCl, 300 mM NaCl, pH 7.4 or PBS, pH 7.4, 8% trehalose.
Note: For SPR assay, please replace the buffer. Primary amine components (e.g., Tris, imidazole) can affect protein-coupled chips.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.