GFP Dipstick Kit (ab119691)

Overview

• Product nameGFP Dipstick Kit
• Precision

  Intra-assay

  Sample	n	Mean	SD	CV%
  1	6			7.5%
• Tests
  1 x 48 test
• Sample type
  Cell culture extracts, Tissue Extracts
• Assay typeSandwich
• Sensitivity
  50 pg/ml
• Range
  0.05 ng/ml - 400 ng/ml
• Recovery

  164 %
  

  Sample specific recovery

  Sample type	Average %	Range
  Cell culture supernatant	164	133% - 200%

• Notes

  Green fluorescence protein (GFP) is a 27 kDa protein derived from the jellyfish Aequorea victoria, which emits green light (emission peak at a wavelenth of 509 nm) when excited by blue light (excitation peak at a wavelenth of 395 nm). GFP has become an invaluable tool in cell biology research, since its intrinsic fluorescence can be visualized in living cells. GFP has been widely used as a reporter for gene expression, enabling researchers to visualize and localize GFP-tagged proteins within living cells without the need for chemical staining.

  While GFP expression can sometimes be detected by fluorescence emission in cells and tissues, determination of very low levels of expression for optimization of expression is difficult and time consuming. This GFP Dipstick ELISA Kit is lateral flow immunoassay developed for an easy, rapid and sensitive detection of GFP or GFP fusion proteins in cells or tissue samples. GFP may be accurately quantified with the use of an immunochromato-reader (MS1000) by comparing the signal from an unknown sample with that of a known recombinant GFP standard curve. The kit has a detection sensitivity limit of 50 pg/mL GFP. Each kit provides sufficient reagents to perform up to 48 assays including standard curve and GFP samples. Note, it is possible that improper folding of GFP or the presence of a fusion partner may prevent GFP binding in this kit.
  
  Storage:Store all components at 4°C. This kit is stable for 6 months from receipt. After reconstitution the standard should be stored at -80°C. 10X blocking, dipsticks and gold-conjugated GAR may be stored at room temperature in the provided containers and out of direct sunlight.

Properties

• Storage instructionsStore at +4°C. Please refer to protocols.

• Components	48 tests
  10X Blocking Buffer	1 x 1ml
  10X Detector Antibody	1 x 140µl
  Dipsticks	1 x 48 tests
  Extraction Buffer	1 x 25ml
  GFP Microplate	2 units
  GFP Standard	1 x 100ng
  Wash Buffer	1 x 5ml

• Research Areas
  • Kits/ Lysates/ Other
  • Fluorescent ligands

  • Tags & Cell Markers
  • Fusion / Marker Proteins
  • GFP

  • Cell Biology
  • Other Antibodies
  • Drugs and Chemicals

GFP Dipstick Kit images

• 

  ELISA - GFP Dipstick (ab119691)
  Figure 1. Example standard curve.
• 

  ELISA - GFP Dipstick (ab119691)
  Figure 2. HeLa cell extract at 1mg/mL were spiked with a titration series of GFP recombinant protein (ab98191) and the dipstick signal intensity was compared with that obtained with GFP recombinant protein diluted in extraction buffer. Results show a near 1:1 fit (R2 = 0.99) and maximum recoverability of GFP signal in a protein extract milieu.
• 

  ELISA - GFP Dipstick (ab119691)
  Figure 3. Instrument-Free visual readout: The calibration curve generated with the standard protein can be made into a “visual reference card”. The test sample is then compared against the visual reference card by matching up the reporter line density of the processed dipstick in the appropriate slot. The value (or value range) can then be interpolated based on the position relative to the antigen levels of the lines immediately to the left and to the right.

Protocols

Protocol Booklet