Active Nerve Growth Factor (NGF)

神经生长因子(NGF)活性蛋白

[PROPERTIES]

Source: Prokaryotic expression. Host: E. coli

Residues: Glu19~Arg239

Tags: N-terminal His-tag

Purity: >98%

Endotoxin Level: <1.0EU per 1μg (determined by the LAL method).
Buffer Formulation: 20mM Tris, 150mM NaCl, pH8.0, containing 0.01% sarcosyl, 5% trehalose. Applications: Cell culture; Activity Assays; In vivo assays.
(May be suitable for use in other assays to be determined by the end user.)

Predicted isoelectric point: 10.1

Predicted Molecular Mass: 29.2kDa

Accurate Molecular Mass: 30&33kDa as determined by SDS-PAGE reducing

conditions. Phenomenon explanation:

The possible reasons that the actual band size differs from the predicted are asfollows:

1. Splice variants: Alternative splicing may create different sized proteins from the same gene.
2. Relative charge: The composition of amino acids may affects the charge of the protein.
3. Post-translational modification: Phosphorylation, glycosylation, methylation etc.
4. Post-translation cleavage: Many proteins are synthesized as pro-proteins, and then cleaved togive the active form.

5. Polymerization of the target protein: Dimerization, multimerization etc.

[USAGE]

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0mg/mL. Do not vortex.

[STORAGE AND STABILITY]

Storage: Avoid repeated freeze/thaw cycles.

Store at 2-8ºC for one month.

Aliquot and store at -80ºC for 12 months.

Stability Test: The thermal stability is described by the loss rate. The loss ratewas determined by accelerated thermal degradation test, that is, incubate theprotein at 37ºC for 48h, and no obvious degradation and precipitation wereobserved.The loss rate is less than 5% within the expiration date underappropriate storage condition

[SEQUENCE]

[ ACTIVITY ] 

Mechanism: Nerve growth factor (NGF) is a neurotrophic factor and neuropeptideprimarily involved in the regulation of growth, maintenance, proliferation, andsurvival of certain target neurons. When the pheochromocytoma cell line PC12 isexposed to nerve growth factor (NGF), the cells respond over a period of a weekby ceasing cell division and extending neurites (Greene and Tischler, 1976). Thecells were grown in Ham's F12K containing 5% fetal calf serum and 10% horseserum on polystyrene tissue culture plates or collagen coated plates. When cellsreached log phase growth and fresh medium was added together with 10ng/mL ofNGF, then cells were observed by inverted microscope everyday.Result 1: The cell division ceased and morphological differentiation of PC12 cellswas observed obviously after incubation with NGF (10ng/mL) for 6 days. Controlgroup which received no NGF displayed no neurite outgrowth and cells multiplyrapidly

NGF is also involved in pathways besides those regulating the life cycle of neurons. Besides, Alpha-2-Macroglobulin (a2M) has been identified as an interactor of NGF,thus a binding ELISA assay was conducted to detect the interaction ofrecombinant human NGF and recombinant human a2M. Briefly, NGF were dilutedserially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μL were thentransferred to a2M-coated microtiter wells and incubated for 2h at 37℃. Wellswere washed with PBST and incubated for 1h with anti-NGF pAb, then aspiratedand washed 3 times. After incubation with HRP labelled secondary antibody, wellswere aspirated and washed 3 times. With the addition of substrate solution, wellswere incubated 15-25 minutes at 37℃. Finally, add 50µL stop solution to the wellsand read at 450nm immediately. The binding activity of NGF and a2M was shownin Figure 2, and this effect was in a dose dependent manner

[IDENTIFICATION]

[IMPORTANT NOTE]

The kit is designed for research use only, we will not be responsible for any issue if the kit wasused in clinical diagnostic or any other procedures.

本产品只适用于科研，不能用于临床诊断。严禁用于临床医疗及其他非科研用途！

以实际收货产品说明书为准，网站说明书仅供参考。