PEPTITE-2000

  • PEPTITE-2000® is a synthetic peptide containing the RGD cell attachment sequence.
  •  It has been designed to bind onto plastic surfaces via a hydrophobic C-terminal sequence.
  •  PEPTITE-2000® can be substituted as the primary cell attachment component in a chemically defined animal component-free culture system.
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General

PEPTITE-2000® is a synthetic peptide containing the RGD cell attachment sequence found in fibronectin, vitronectin and many other matrix and serum proteins.   Additionally, it has been designed to bind onto plastic surfaces via a hydrophobic C-terminal sequence. PEPTITE-2000® can be substituted as the primary cell attachment component in a chemically defined animal component-free culture system.  PEPTITE 2000® is supplied in sterile 5 mg quantities.

The tripeptide Arg-Gly-Asp (RGD) is an important protein sequence in the binding of proteins to cell surfaces. The RGD motif was initially identified in fibronectin as the minimal sequence that mediates cell attachment. Research has also shown that the RGD motif in present in other proteins such as vitronectin, osteopontin, collagens, thrombospondin, fibrinogen and many other factors. Many adhesive cell surface receptors recognize the RGD sequence for cell attachment.

RGD PEPTITE-2000® has been specifically developed for ease-of-use for the researcher. This product is provided in a 5 mg package size to provide sufficient quantity of product for coating a large surface area. The product has been sterilized and is ready-to-use after proper dilution.

Application

PEPTITE-2000® is used to coat tissue culture plasticware for enhanced cell attachment and adhesion. The optimal concentration for cell attachment and culture may differ for various cell types. Some experimentation may be required to determine the optimal conditions for individual cell culture systems. A typical working concentration may range from 0.1 to 10 μg/ml. PepTite-2000 is not for human use as supplied.

Composition

PEPTITE-2000® Characteristics

Parameter, Testing, and Method
PEPTITE-2000®
Part Number  5020

Form

Powder

Package Size

5 mg

Sterilization

Irradiation Sterilized

Peptide Sequence

Ac-GrGDIPASSKGGGGrLLLLLLr-NH2

Purity Confirmed by
RP-HPLC

>95%

Identity Confirmed by
Amino Acid Analysis

Characteristic

Identity Confirmed by
Mass Spectrometry

Characteristic

Peptide Content Confirmed by
Nitrogen Analysis

Characteristic
Sterility
No Growth
Endotoxin (LAL)
< 0.5 EUmL

Storage Temperature

Desiccated, 2-10 °C

Data Sheets

Printable PDF Version

For research use only

PRODUCT DESCRIPTION

PEPTITE-2000® has been specifically developed for ease-of-use for the researcher. This product is provided in a 5 mg package size to provide sufficient quantity of product for coating a large surface area. The product has been sterilized and is ready-to-use after proper dilution.

STORAGE

It is recommended that PEPTITE-2000® be stored desiccated at 2 to 10°C or below.

CHARACTERIZATION

  • Purity: PEPTITE-2000® has a purity of >95% based on HPLC analysis.
  • Quantity: The quantity of peptide in each container is 5 mg.
  • Sterilization: PEPTITE-2000® has been sterilized by Ebeam irradiation.
  • Peptide Sequence: Ac-G(d)RGDIPASSKGGGGS(d)RLLLLLL(d)R-Amide

INSTRUCTIONS FOR USE

Use these recommendations as guidelines to determine the optimal coating conditions for your culture system. To maintain sterility, perform all operations in a laminar flow hood. Two options are provided:

Procedure A
  1. Remove cap and add 5 ml of serum-free medium or PBS to the bottle.
  2. Replace cap and vortex contents vigorously. Ensure that the PEPTITE-2000® is completely solubilized. The solution will remain slightly hazy.
  3. Transfer desired volume of solution from the bottle to a dilution vessel. Dilute to desired concentration using serum-free medium or PBS. A typical working concentration may range from 0.1 to 10 μg/ml.
  4. Add appropriate amount of diluted material to culture surface.
  5. Incubate at room temperature or 37°C, covered, for 1-2 hours.
  6. After incubation, aspirate remaining material.
  7. Rinse plates carefully with dH2O– avoid scratching bottom surface of plates.
  8. Plates are ready for use. They may also be stored at 2-8°C damp or air dried if sterility is maintained.
  9. Store remaining solubilized PEPTITE-2000® at 2 to 10°C.

Additional note: Include divalent cations (Calcium, Magnesium, or Manganese) in cell attachment solution to obtain optimum cell binding.

Procedure B
  1. Remove cap and add 5 ml of sterile 70% Ethanol
  2. Replace cap and vortex contents. Ensure that the PEPTITE-2000® is completely solubilized.
  3. Transfer desired volume of solution from the bottle to a dilution vessel. Dilute to the desired concentration using 70% Ethanol. Concentrations from 0.1 to 10 ug/ml should be tested.
  4. Add appropriate amount of diluted material to culture surface.
  5. Leave the coated container, uncovered, in a laminar flow hood until the wells are dry.
  6. Rinse plates carefully with dH2O– avoid scratching bottom surface of plates.
  7. Plates are ready for use.
  8. Store remaining solubilized PEPTITE-2000® at 2-10°C.

PRECAUTION

Follow typical laboratory safety practices when handling PEPTITE-2000®

References