• May 29, 2021

RETRO-TEK HIV-1 P24 Antigen ELISA,

Expected Use: The RETRO-TEK HIV-1 p24 Antigen ELISA 2.0 is an enzyme-linked immunoassay used to detect human immunodeficiency P24 antigen of virus type 1 (HIV-1) in cell culture media. It can be used to monitor the purification and biochemical behavior of HIV-1 or determine the titer of HIV-1-based lentiviral samples. For various HIV-1 isolates, this assay detects p24 from HIV-1 clades A-F.

Principle Of The Test: The wells of the microplate are coated with a monoclonal antibody specific for the HIV-1 p24 gag gene product. HIV-1 p24 antigen in the sample is specifically captured on the immobilized antibody during the incubation of the sample. The captured antigen is reacted with horseradish peroxidase (HRP) conjugated human anti-HIV-1 antibody. After the subsequent addition of substrate, the color develops as the HRP enzyme reacts.

The resulting optical density is proportional to the amount of HIV-1 p24 antigen present in the sample. The absorbance values ​​of a set of standard dilutions are then plotted. The amount of p24 is determined by interpolation from a point-to-point graph or linear regression analysis of the standard curve.

REAGENTS

Supplied materials:

  • Microplate coated with p24 HIV-1 antibody for 96 determinations, 5 plates: 12×8-well strips. Wells covered with a murine
    Monoclonal antibody against HIV-1 p24.
  • HIV-1 Detector Antibody p24, 5 x 12 ml: Contains human antibody to HIV-1 conjugated to HRP (horseradish peroxidase)
    purified from HIV-1 human source material.
  • HIV-1 p24 antigen standard, 5 x 0.5 ml: Contains HIV-1 IIIB detergent interrupted heat-inactivated viral antigen, goat
    serum, Triton X-100 and sodium azide.
  • Lysis buffer, 5 x 5 ml: Triton X-100 in PBS and 2-chloroacetamide.
  • Assay Diluent, 5 x 100 ml: Contains goat serum, PBS, Triton X-100, and 2-chloroacetamide.
  • 10X Plate Wash Buffer, 5 x 125 ml: Contains PBS, Tween 20, and 2-chloroacetamide.
  • Substrate, 5 x 12 ml: Contains tetramethylbenzidine (TMB).
  • Microplate sealers (1 pack): 10 sealers per pack
  • Stop Solution, 5 x 12 ml: Contains hydrochloric acid (HCl).
  • Plate Sealants: 5 x10
  • Resealable plastic bag: 5

Storage: Store all kit reagents at 2 ° – 8 ° C. DO NOT FREEZE. When properly stored, the kit is stable until the date indicated on the box.

Materials Required But Not Provided:

  • Disposable gloves
  •  Adjustable, validated, mono and multichannel pipettes
  • Test tubes and racks to prepare samples and control dilutions.
  • Graduated cylinders and assorted beakers
  • Validated automatic microplate washer or manual vacuum aspiration equipment
  • Incubator validated for 37 ° C ± 1 ° C
  • Validated microplate reader
  • Timer
  • 1% sodium hypochlorite as a disinfectant. It can be prepared with homemade bleach.
  • Distilled or deionized water

Precautions

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC USE in vitro.

  • Before performing the test read all instructions carefully.
  • Use universal precautions when handling kit components and test samples.
  • To avoid cross-contamination, use separate pipette tips for each sample.
  • When testing potentially infectious human specimens, comply with all applicable federal, state, and local regulations.
    regarding the disposal of biohazard materials.
  • The HIV-1 p24 antigen standard contains sodium azide as a preservative. Sodium azide can react with lead or copper pipes.
    to form explosive metal azides. Flush pipes with large amounts of water when disposing to avoid the accumulation of azides in drains.
  • The stop solution contains hydrochloric acid, which can cause severe burns. In case of contact with eyes or skin, rinse.
    immediately with water and seek medical assistance. Wear protective clothing and goggles.
  • The material of human origin used in the manufacture of the HIV-1 Detector Antibody has been tested and found negative for
    Hepatitis B surface antigen. The viral lysate used to prepare the HIV-1 antigen p24 standard has been inactivated by
    chemical disruption and heating. Handle these reagents as if they are capable of transmitting infectious agents.
  • Although the p24 antigen is well conserved, some HIV-1 isolates and recombinant clones may show a mildly reactive reaction.

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