Certificate of Analysis and Data Sheet

Source:
E.Coli

Catalog No.
CYT-331

Background:

This ligand binds to Fms-Like Tyrosine Kinase-3 (FLT-3).Human FLT3 ligand also stimulates the proliferation of cells expressing murine FLT-3 receptors. The effects of FLT-3 ligand are synergized by coexpression of G-CSF , GM-CSF , M-CSF , IL3 and SCF . FLT-3 ligand promotes long-term expansion and differentiation of human pro-B-cells in the presence of IL7 or in combination of IL7 and IL3 .

Description :

Recombinant Human Flt3-Ligand produced in E.Coli is non-glycosylated, polypeptide chain containing 155 amino acids and having a molecular mass of 17605 Dalton.
Recombinant FLT3 Ligand is purified by proprietary chromatographic techniques.

Physical Appearance:

Sterile Filtered White lyophilized (freeze-dried) powder.

Formulation:

FLT-3 was lyophilized with no additives.

Solubility:

It is recommended to reconstitute the lyophilized FLT-3 in sterile 18MΩ-cm H2O not less than 100ug/ml, which can then be further diluted to other aqueous solutions.

Stability:

Lyophilized FLT3 although stable at room temperature for 3 weeks, should be stored desiccated below -18 C. Upon reconstitution FLT3 should be stored at 4 C between 2-7 days and for future use below -18 C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please avoid freeze-thaw cycles.

Purity:

Greater than 98.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.

Amino acid sequence:

The sequence of the first five N-terminal amino acids was determined and was found to be Thr-Gln-Asp-Cys-Ser.

Dimers and aggregates:

Less than 1% as determined by silver-stained SDS-PAGE gel analysis.

Biological Activity:

ProSpec旧 rHuFlt3-Ligand is fully biologically active when compared to standard. The ED50, calculated by the dose-dependant stimulation of the proliferation of human AML5 cells is less then 1.0 ng/ml, corresponding to a specific activity greater then 1 x 10⁶ units/mg.

Endotoxin:

Less than 0.1 ng/ug (IEU/ug) of FLT-3.

Protein content:

Protein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 .
2. Analysis by RP-HPLC, using a standard solution of FLT3 as a Reference Standard.

Usage:

This material is offered by ProSpec-TechnoGene for research, laboratory or further evaluation purposes.

Gene:

Name:FLT3LG

Protein synonyms/aliases:

SL cytokine precursor (Fms-related tyrosine kinase 3 ligand)
    (Flt3ligand)
    (Flt3L).

Protein Family:

Protein Domains:

Domains:
•IPR004213   Flt3 ligand

Protein links:

Precursor- Protein structure and amino acid sequence:

Latest Publications:

1. Significant reduction of laparotomy-associated lung metastases and subcutaneous tumors after perioperative immunomodulation with flt3 ligand in mice.
Surg Innov 2005 Dec;12(4):319-25
2. Kinetics of plasma FLT3 ligand concentration in hematopoietic stem cell transplanted patients.
Leuk Lymphoma 2006 Jan;47(1):77-80
3. Cancer gene therapy using in vivo electroporation of Flt3-ligand.
Int J Oncol 2005 Aug;27(2):457-63
4. Flt3 ligand-generated murine plasmacytoid and conventional dendritic cells differ in their capacity to prime naive CD8 T cells and to generate memory cells in vivo.
J Immunol 2005 Jul 1;175(1):189-95
5. Mesenchymal stem cells support expansion of in vitro irradiated CD34(+) cells in the presence of SCF, FLT3 ligand, TPO and IL3: potential application to autologous cell therapy in accidentally irradiated victims.
Radiat Res 2005 Jul;164(1):1-9
6. Functional expression, purification, and characterization of human Flt3 ligand in the Pichia pastoris system.
Protein Expr Purif 2005 Aug;42(2):246-54

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中華民國95年06月06日更新