FaP-Kit (Fix & Perm)

Details zu Produkt Nr. ABIN5662622, Anbieter: Anmelden zum Anzeigen
Applikation
Sample Preparation (Prep)
Optionen
Hersteller
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Verwendungszweck FIXATION, PERMEABILIZATION and RED BLOOD CELL Lysis in a 2-step-method which enables detection of intracellular antigens for following flow cytometric analysis of whole blood and other samples
Spezifität Fix and Perm, Fix & Perm, Fixation, Permeabilization, Intacellular Staining, RBC-Lyse, Lysing solution, Lysing buffer
Produktmerkmale FaP-Kit contains appropriate ready-to-use reagents sufficient for mild fixation and inducing of permeability in cytoplasmatic membranes of leucocytes and immunologic detection of antigens by means of antibodies and subsequent analysis by flow cytometry or microscopy.
Applikationshinweise 1 test = 100 μL FIX + 100 μL PERM (200 tests = 2x 20ml)
Kommentare

- Fast and efficient - whole procedure can be performed in one hour
- Enables simultaneous detection of both intracellular and cell surface markers
- Softly fixed cells with conserved flow cytometric scatter properties
- Specific formulation for reducing background
- Quality control assures minimization of lot-to-lot variations

Probenmenge 100 μL
Protokoll In a first step the cells are fixed by using FIX reagent. Subsequently the cells are permeabilized and erythrocytes are lysed by using PERM reagent. Intracellular antigen detection goes on after permeabilization step. Surface marker detection goes on prior to the fixation step or parallel to the intracellular detection step provided that antibodies against surface markers are able to detect fixed surface markers as well.
Testdurchführung
  • Add 100 μL of the sample (blood, bone marrow sample or other cell suspension) into your test tube (≥ 5ml). The sample must be completely on the bottom of the tube.
    - For immuno-staining add a sufficient volume (usually 1-20 μL depending on the manufacterer) of antibody solution into the sample, vortex and incubate
    - Incubate 15 min / RT
    - Add 100 μL of FIX reagent to the test tube, vortex and incubate. Several short vortex cycles within the incubation may optimize the results.
    - Incubate 15 min / RT
    - Add 4 mL PBS and centrifuge for 5 min at 350g and remove completely the supernatant by aspiration
    - Add 100 μL of PERM reagent to the test tube, shake gently approx. 5 sec and incubate.
    - Incubate 5 min / RT
    - For intracellular immuno-staining add a sufficient volume (usually 1-20 μL depending on the manufacterer) of antibody solution into the sample, vortex and incubate. Make sure that the cell pellet is completely resuspended. Several short vortex cycles within the incubation may optimize the results.
    - Incubate 15 min / RT
    - Add 4 mL PBS and centrifuge for 5 min at 350g and remove completely the supernatant by aspiration
    - Resuspend the pellet in approx. 0.5 mL PBS for direct analysis or in 0.5 mL 0.1 % formaldehyd in PBS for analysis within 24h
Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Buffer FaP-KIT contains fomaldehyde
Lagerung RT
Informationen zur Lagerung Store at 10-25°C
Closed vials: 3 years after delivery
Opened vials: 6 months
Bilder des Herstellers
Flow Cytometry (FACS) image for FaP-Kit (Fix & Perm) (ABIN5662622) Example: Intracellular detection of granzyme A, granzyme B and perforin in whole bloo...