Product Description |
GenScript’s
Lentivirus Titer p24 ELISA kit (Cat. No. L00938) is a Sandwich ELISA designed for quantitatively
measuring the physical titer of any HIV-1 based lentivirus in
cell culture supernatant. For lentivirus purification and production,
lentivirus titration is an essential characterization procedure for routine
quality control and lot release, as well as for purification process
optimization. This kit makes lentivirus titration easy,
accurate and reproducible. Lentivirus vector is
derived from human immunodeficiency virus-1 (HIV-1). The vector, which contains
a pseudotyped envelope with vesicular stomatitis virus envelope G (VSV-G)
protein, has the ability to transfer gene(s) of interest into dividing and
non-dividing cells. Many studies show that Lentiviral vectors have successfully
delivered genes into neurons, lymphocytes, and macrophages. The vector is
proven to be effective in transducing brain, liver, muscle, and retina in vivo without toxicity or immune
responses. The vector is also used to integrate siRNA efficiently in a wide
variety of cell lines. |
Linearity Range |
31.25 pg/mL - 2000 pg/mL |
Sensitivity |
LoB: 14.13 pg/mL LoD: 29.42 pg/mL LoQ: 39.03 pg/mL |
Precision |
Intra-assay: CV ≤ 10%; Inter-assay: CV ≤ 15% |
Assay Temperature |
25°C |
Assay Time |
1 hour 40 minutes |
Kit Contents |
Component |
Quantity/Size |
Part No. |
Capture Plate |
1 plate |
E1-80 |
Standard Stock |
1 vial (100 μL) |
E1-10 |
Lysis Solution |
1 vial (1.6 mL) |
E1-A0 |
Biotin Anti-p24 Antibody |
1 bottle (12 mL) |
E1-20 |
Streptavidin-HRP |
1 bottle (12 mL) |
E1-30 |
TMB Solution |
1 bottle (12 mL) |
A1-40 |
20× Wash Solution |
1 bottle (40 mL) |
A1-70 |
Stop Solution |
1 bottle (6 mL) |
A1-50 |
Plate Sealer |
2 pieces |
N/A |
|
Storage |
The unopened kit is stable for at least 12 months from the date of
manufacture at 2°C to 8°C, and the opened kit is stable for up to 1 month from
the date of opening at 2°C to 8°C. |
Assay Principle |
Lentivirus Titer p24 ELISA kit utilizes two HIV-1 p24 monoclonal antibodies which bind to different epitopes of the p24 protein. When standards or samples are added to the capture plate, the p24 protein in the sample can be captured on the plate coated with the HIV-1 p24 capture antibody. Then the HIV-1 p24 detection antibody conjugated with biotin is added to interact with the p24 protein bound on the plate. Streptavidin-Horseradish Peroxidase conjugate (Streptavidin-HRP) is added to interact with the biotin conjugated anti-p24 antibody (Biotin Anti-p24 Antibody). After washing steps, 3,3',5,5'-Tetramethylbenzidine solution (TMB Solution) is added resulting in formation of blue color. The reaction is stopped by adding Stop Solution. Application of the Stop Solution results in the color changing from blue to yellow. The intensity of the color can be read at 450 nm by a microplate reader.
The quantity of p24 protein in the sample is precisely quantified against a p24 standard curve. p24 values are then correlated to the lentivirus titer of packaging cell culture supernatants.
Figure 1: Schematic diagram of Lentivirus Titer p24 ELISA kit |
Schematic diagram of Lentivirus Titer p24 ELISA kit
For
laboratory research use only. Direct human use, including taking orally and
injection and clinical use are forbidden.