Perforin Human ProcartaPlex™ Simplex Kit
Perforin Human ProcartaPlex™ Simplex Kit
Citas y referencias (2)
Invitrogen™

Perforin Human ProcartaPlex™ Simplex Kit

El kit de perforina humana Simplex ProcartaPlex mide la proteína perforina y está diseñado para combinarse con otros kits SimplexMás información
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Número de catálogoCantidad
EPX010-12306-90196 pruebas
Número de catálogo EPX010-12306-901
Precio (CLP)
-
Cantidad:
96 pruebas
El kit de perforina humana Simplex ProcartaPlex mide la proteína perforina y está diseñado para combinarse con otros kits Simplex para que pueda crear su propio panel multiplex que utilice tecnología Luminex xMAP para la detección o cuantificación de proteínas. Cuando se combinan varios kits Simplex (es decir, cuando no se utiliza un panel multiplex preconfigurado), solo se necesita un kit de tampón (se vende por separado) para cada placa de ensayo, independientemente del tamaño del complejo.

Acerca de los ensayos ProcartaPlex para la plataforma Luminex

Los inmunoensayos ProcartaPlex se basan en los principios de ELISA tipo sándwich y utilizan dos anticuerpos altamente específicos que se enlazan a diferentes epítopos de una proteína para cuantificar todos los objetivos de proteína simultáneamente utilizando un instrumento Luminex. Los ensayos multiplex ProcartaPlex requieren tan solo 25 µl de plasma o suero o 50 µl de sobrenadante de cultivo celular y solo cuatro horas para obtener los resultados analizados.

• Más resultados por muestra: mida múltiples objetivos de proteínas simultáneamente en una única muestra de 25–50 µl

•Tecnología Luminex consolidada: plataforma de multiplexing de referencia para detección y cuantificación de proteínas

Los ensayos ProcartaPlex utilizan tecnología Luminex Xmap (perfil multianalito) para la detección y cuantificación simultáneas de hasta 65 proteínas diana en una sola muestra de 25–50 µl de plasma, suero, sobrenadantes de cultivo celular y otros fluidos corporales.

Los gránulos Luminex del ensayo ProcartaPlex se tiñen internamente con proporciones precisas de fluoróforos rojos e infrarrojos para crear firmas específicas que pueden ser identificadas por los sistemas de detección Luminex Xmap (por ejemplo, Luminex 200, FLEXMAP 3D y MAGPIX). Similar a un ELISA tipo sándwich, el ensayo ProcartaPlex utiliza pares de anticuerpos emparejados para identificar la proteína de interés. En un ensayo multiplex, cada gránulo espectralmente único se etiqueta con anticuerpos específicos para una sola proteína diana, y las proteínas ligadas se identifican con anticuerpos biotinilados y estreptavidina-R-ficoeritrina (RPE). La conjugación de anticuerpos específicos de proteínas con un gránulo distinto permite el análisis de varios analitos en un solo pocillo.

La diferencia más significativa entre un ensayo ProcartaPlex y uno ELISA es que el anticuerpo de captura del ensayo ProcartaPlex se conjuga con un gránulo y no se absorbe en el pocillo de la microplaca, por lo que los reactivos del ensayo ProcartaPlex flotan en la solución. Para la detección, el instrumento Luminex 200, por ejemplo, contiene dos láseres, uno para distinguir la firma espectral de cada gránulo y el segundo para cuantificar la cantidad de fluorescencia RPE, que es proporcional a la cantidad de proteína presente en la muestra. Los ensayos multiplex ProcartaPlex pueden perfilar más proteínas diana utilizando mucha menos muestra en el mismo tiempo que se tarda en realizar un ELISA tipo sándwich tradicional.

Los paneles multiplex ProcartaPlex están disponibles en múltiples formatos en seis especies (humano, ratón, rata, primate no humano, porcino y canino).
Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.
Especificaciones
Intervalo del ensayoSegún lo determinado para el lote 1: De 73,24 a 30 000 pg/ml
Sensibilidad del ensayoSegún lo determinado para el lote 1: 158,4 pg
Tipo de esferaPerforina [53 ]
Para utilizar con (equipo)Instrumentos Luminex™
FormatoKit Simplex
génicaperforina 1
Alias de genP1; PFP; HPLH2
ID de gen (Entrez)5551
Símbolo de genPRF1
Línea de productosProcartaPlex
ExpresiónPerforina-1
Subtipo de proteínacitolisina, proteína formadora de poros de linfocitos, perforina 1 (proteína formadora de poros)
Tipo de muestraSobrenadantes de cultivo celular, suero y plasma, Plasma, Cell Culture Supernatants
Volumen de muestraSuero, plasma: 25 μl; CCS: 50 μl
Condiciones de envíoHielo húmedo
ID. de UniProtP14222
CombinabilityCombinable
Tipo de productoKit Simplex
Cantidad96 pruebas
Research AreaImmunology, Oncology
EspecieHumano
Unit SizeEach
Contenido y almacenamiento
  • 1 vial de gránulos de captura (50X)
  • 1 vial de anticuerpos de detección biotinilada (50X)
  • 2 viales de perforina estándar humana (liofilizada)
  • Almacenar entre 2 ° y 8 °C.

Preguntas frecuentes

What is the size of the Luminex beads you currently use?

The beads used in our Luminex instrument-compatible ProcartaPlex and QuantiGene Plex assays are 6.5 micron superparamagnetic beads.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

I am interested in performing Luminex assays using BioSource kits, and I have a Luminex xMAP system. Besides the kits and system, what other reagents and equipment will I need?

The following is a list of general lab supplies that are required for running BioSource immunoassays on the Luminex xMAP system:
1) Sonicating water bath
2) Orbital shaker
3) Vortexer
4) Repeating and/or multi-channel pipetter (not required, but recommended)
5) Calibrated adjustable precision pipettes, with disposable plastic tips
6) Glass/plastic tubes and racks for preparing reagents
7) Graduated cylinder and container for preparing wash solution
8) Aluminum foil
9) Deionized or distilled water.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Do the Luminex beads require special care in handling?

The Luminex beads should be protected from light because they are susceptible to photobleaching. We recommend protecting the beads by keeping containers covered with aluminum foil during all incubation steps, and exercising care during handling. The beads should not be frozen, subjected to excessive heat, or exposed to organic solvents.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Why would the Luminex acquisition software display "Sample Empty" messages during analysis?

(1) The user did not properly aliquot the diluted beads, such that no beads were actually added to the wells (make sure that the bead concentrates are sonicated and vortexed well, then check the pipet tip to ensure that air bubbles were not drawn up)
(2) The user missed loading diluted beads to some wells, which is likely since the small volume is clear and difficult to visualize in the clear plastic plate (we have now addressed this customer difficulty by coloring each of the Buffer Reagent Kit components)
(3) The user applied too much vacuum pressure at some point during the wash steps, or allowed the pressure to spike even once, such that the filter membrane tore in a few wells releasing the beads (make sure that the vacuum manifold pressure is kept below 5mm/in Hg, depending on their system -- a good rule of thumb is that it should take a full 3-second count to GENTLY empty the wells of 200uL)
(4) The user did not properly sonicate and vortex the beads prior to dilution, such that the percent of bead aggregation was high and the instrument was unable to find enough single beads to meet the events/bead value designated by the customer (make sure that the Bead Concentrate tube is put into the waterbath all the way to the cap, since the tube is hollow until the top third)
(5) The user lost beads by shaking the plate too aggressively or handling it improperly (make sure that the orbital shaker is set to a speed that allows for maximum vortex in the wells without spillage)
(6) The user exposed the beads to an excess of light during storage or running of the assay, such that some but not all of the beads were photobleached and therefore falling outside the acceptable range for each bead region (make sure that the plate is covered on the top/sides with foil throughout the assay, away from Windows and spotlights, and that the bead component of the kits is stored in the dark)*
(7) There was a clog in the sample needle, such that the instrument was unable to take up enough sample to meet the number of events requested per bead region (suggest that the user follow the manual instructions for dislodging a clog, which include several Back Flush steps and may require removal of the needle for sonication with probe alignment).

* Some of the older Antibody Bead Kits still have clear plastic tops instead of black ones. In cases where customers store kits in lit refrigerators, or keep them open on the lab bench, even a few hours of light exposure is enough to photobleach beads. It is important to note, in general, that higher number bead regions are more susceptible to photobleaching. In order to draw conclusions about the source of the difficulty, we would ask to see the data, specifically the Masterplex QT file, which would enable us to examine the pattern of "Sample Empty" occurrences in addition to the bead counts per well.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

What are the Luminex beads made of?

The beads are made of polystyrene.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

View all Perforin Human ProcartaPlex™ Simplex Kit FAQs

Citations & References (2)

Citations & References
Abstract
HTLV-2 Enhances CD8+ T Cell-Mediated HIV-1 Inhibition and Reduces HIV-1 Integrated Proviral Load in People Living with HIV-1
Authors:Abad-Fernández, M. et al
Journal:Viruses
PubMed ID:36366570
'People living with HIV-1 and HTLV-2 concomitantly show slower CD4+ T cell depletion and AIDS progression, more frequency of the natural control of HIV-1, and lower mortality rates. A similar beneficial effect of this infection has been reported on HCV coinfection reducing transaminases, increasing the spontaneous clearance of HCV infection ... More
HTLV-2 Enhances CD8(+) T Cell-Mediated HIV-1 Inhibition and Reduces HIV-1 Integrated Proviral Load in People Living with HIV-1.
Authors:Abad-Fernández M,Hernández-Walias FJ,Ruiz de León MJ,Vivancos MJ,Pérez-Elías MJ,Moreno A,Casado JL,Quereda C,Dronda F,Moreno S,Vallejo A
Journal:Viruses
PubMed ID:36366570
People living with HIV-1 and HTLV-2 concomitantly show slower CD4(+) T cell depletion and AIDS progression, more frequency of the natural control of HIV-1, and lower mortality rates. A similar beneficial effect of this infection has been reported on HCV coinfection reducing transaminases, increasing the spontaneous clearance of HCV infection ... More