///Rat Ciliary Neurotrophic Factor Recombinant

Rat Ciliary Neurotrophic Factor Recombinant

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$70.00$2,700.00

SKU: RKP20294 Tags: ,

Description

Accession
P20294
Source
Optimized DNA sequence encoding Rat Ciliary Neurotrophic Factor mature chain was expressed in Escherichia Coli.
Molecular weight
Native rat CNTF has a calculated molecular mass of approximately 22 kDa. Recombinant CNTF is a monomer protein consisting of 200 amino acid residue subunits, and migrates as an approximately 22 kDa protein under non-reducing and reducing conditions in SDS-PAGE.
Purity
>98%, as determined by SDS-PAGE and HPLC
Biological Activity
The ED(50) was determined by the dose-dependent proliferation of Human TF1 cells was found to be in the range of 1 ng/ml.

Protein Sequence
MAFTEHSPLT PHRRDLCSRS IWLARKIRSD LTALTESYVK HQGLNKNINL DSADGMPVAS TDQWSELTEA ERLQENLQAY RTFHVLLARL LEDQQVHFTP TEGDFHQAIH TLLLQVAAFA YQIEELMILL EYKIPRNEAD GMPINVGDGG LFEKKLWGLK VLQELSQWTV RSIHDLRFIS SHQTGIPARG SHYIANNKKM
Endotoxin
Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than.1 ng/µg(1EU/µg).
Presentation
Recombinant Rat Ciliary Neurotrophic Factor was lyophilized from a.2 μm filtered PBS pH.5.
Reconstitution
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than.1 mg/mL. This solution can then be diluted into other buffers.
Storage
The lyophilized protein is stable for at least years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at° -° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage
This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.
Biological Process
Biological Process
Molecular function
Molecular function

Methods

Preparation of Primary Hippocampal Cultures

  • Primary hippocampal cultures were prepared from wild-type neonatal (E19) rat embryos (timed pregnant Sprague Dawley rats were obtained from Charles River Laboratories, Wilmington, MA) as described previously

RGC survival under normoxia and hypoxia Survival of RGC under normoxia and hypoxia (0.2% O2).

  • RGC survival (ratio of living cell vs. total cells) was measured after treatment with 0, 1, 5 and 25 ng/ml PEDF or 10 ng/ml CNTF (n = 3; treated versus normoxic or hypoxic PEDF-free cultures, *P < 0.05, **P < 0.01; control hypoxic versus normoxic PEDF-free control culture, **°°P < 0.01).

Immunoisolation and Culture of RGC from Postnatal Mouse Retinae

  • Seven days old mice were killed according to institutional guidelines.
  • To isolate RGC, retinae were dissected and incubated for 45 min at 37°C in Dulbecco’s phosphate buffered saline (D-PBS; containing 160 U/ml papain, 200 U/ml DNAse.
  • The tissues were then sequentially triturated in D-PBS containing 0.15% trypsin inhibitor , 650 U/ml DNAse and 1:75 rabbit anti-rat macrophage antibody.
  • Cells were spun down (800′g for 13 min), resuspended in 1% trypsin inhibitor in D-PBS, spun down again and then resuspended in D-PBS containing 0.02% bovine serum-albumin (fraction V).
  • Cell suspension was filtered through a nylon-mesh ( 20 , , , ) and sequentially added to immunopanning plates.
  • Briefly, immunopanning was performed using two subtraction plates (150-mm diameter petri dishes , , ) coated with goat anti-rabbit-IgG ( , , ; 10 μg/ml) to remove microglial cells after incubation of the supernatant with anti-macrophage antibody (WAK Chemie, ).
  • Positive cell isolation was then achieved by incubating the filtrated…

Knockdown of AMIGO3 promote RGC neurite outgrowth in the presence of inhibitory concentration of CME.

  • Representative photomicrographs of βIII-tubulin+ RGC neurite outgrowth in CNTF and siRNA transfected retinal cells in the presence and absence of CME with or without CNTF.
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