Human FASL / CD95LG / CD95 Ligand PicoKine™ Fast ELISA Kit

SKU FEK0337
Size 96wells/kit, with removable strips.
Reactivity Human
Sample Type cell culture supernates, serum and plasma(heparin, EDTA, citrate).
Sensitivity <2pg/ml
Assay Range 15.6pg/ml-1000pg/ml

Introduction

Boster's Picokine™ Fast ELISA kits features the high sensitivity of Picokine™ ELISA kits and the significantly reduced assay run time. Now you can run the entire ELISA assay in less than 1.5 hours.

Compared to regular Picokine ELISA kits, the following sections in the datasheet are different:
1. kit components;
2. preparation before experiment;
3. assay protocol

Overview

Product Name Human FASL / CD95LG / CD95 Ligand PicoKine™ Fast ELISA Kit
SKU/Catalog Number FEK0337
Storage & Handling Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)
Size 96wells/kit, with removable strips.
Description The Fast version of Picokine ELISA kits, assay takes less than 1.5 hours. Detect Human Fas Ligand/FASLG with <2pg/ml sensitivity. Format: 96-well plate with removable strips. Compatible samples: cell culture supernates, serum and plasma(heparin, EDTA, citrate). This is a TMB colorimetric sandwich ELISA kit with short assay time and fast experiment set up. Fas Ligand/FASLG tissue specificity:
Cite This Product Human FASL / CD95LG / CD95 Ligand PicoKine™ Fast ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # FEK0337)
Sample Type cell culture supernates, serum and plasma(heparin, EDTA, citrate).. Anticoagulant(s): heparin, EDTA or citrate
*The recommended anticoagulants are proven to not block the antibody binding sites on the target antigen. Please do not collect blood sample with other anticoagulants that are not specified above or contact us to check for feasibility.
Sensitivity <2pg/ml
Assay Range 15.6pg/ml-1000pg/ml
Immunogen Expression system for standard: CHO; Immunogen sequence: P134-L281
Reactivity Human
Cross Reactivity There is no detectable cross-reactivity with other relevant proteins.
Antibody Clonalities Capture Antibody | Detection Antibody:
monoclonal antibody from mouse|polyclonal antibody from goat

Assay Details

Kit Components

Catalog numberDescriptionQuantity
FEK0337-CAP96-well plate precoated with anti-Human FASLG antibody1
FEK0337-STlyophilized recombinant Human FASLG standard10ng/tube
FEK0337-DAbiotinylated anti-Human FASLG antibody130ul
AR1103Avidin-Biotin-Peroxidase Complex(ABC)400ul
AR1106-1sample diluent buffer30ml
AR1106-2antibody diluent buffer12ml
AR1106-3ABC diluent buffer12ml
AR1104TMB color developing agent10ml
AR1105TMB stop solution10ml
PLA-SEAAdhesive cover4

*Why there is no wash buffer? Our Avidin-Biotin-Peroxidase Diluent contains the detergent (TWEEN) normally present in other companies' ELISA kits. This saves you the step of having to wash with the special wash buffer and achieve similar or better signal to noise ratio. The wash can use regular wash buffers (PBS, TBS etc.) commonly found in labs.

Materials Required But Not Provided

  • Microplate reader in standard size.
  • Automated plate washer.
  • Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
  • Clean tubes and Eppendorf tubes.
  • Washing buffer (neutral PBS or TBS).
  • Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g NaCl; 450μl of purified acetic acid or 700μl of concentrated hydrochloric acid to 1000ml H2
  • Preparation of 0.01 M PBS: Add 8.5g sodium chloride, 1.4g Na O and adjust pH to 7.2-7.

Typical Data Obtained from Human FASL / CD95LG / CD95 Ligand PicoKine™ Fast ELISA Kit

(TMB reaction incubate at 37°C for 15-20min)

Concentration(pg/ml)015.631.262.51252505001000
O.D.0.0080.1140.1960.3790.7401.3712.1212.693

Intra/Inter Assay Precision

Intra-Assay PrecisionInter-Assay Precision
Sample123123
n161616242424
Mean(pg/ml)3618643734189419
Standard deviation2.199.6728.842.112.4733.1
CV(%)6.1%5.2%6.6%6.2%6.6%7.9%

Reproducibility

Three samples with differing target protein concentrations were assayed using four different lots to measure the CV% lot to lot variance.

Reproducibility

To assay reproducibility, three samples with differing target protein concentrations were assayed using four different lots.

LotsLot1 (pg/ml)Lot2 (pg/ml)Lot3 (pg/ml)Lot4 (pg/ml)Mean (pg/ml)Standard DeviationCV (%)
Sample 136414134383.088.1%
Sample 21861971921981934.762.4%
Sample 343749544650747130.216.4%
*number of samples for each test n=16.

*The typical data is obtained from Boster's internal QC result and for reference only. It may differ from the lab test results of the end users. It is more important that the user's lab test results reflect the same linearity demonstrated in the typical data than achieving exactly the same O.D. values.

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id P48023
Gene Name FASLG
Protein Name Tumor necrosis factor ligand superfamily member 6
Alternative Names Tumor necrosis factor ligand superfamily member 6;Apoptosis antigen ligand;APTL;CD95 ligand;CD95-L;Fas antigen ligand;Fas ligand;FasL;CD178;Tumor necrosis factor ligand superfamily member 6, membrane form;Tumor necrosis factor ligand superfamily member 6, soluble form;Receptor-binding FasL ectodomain;Soluble Fas ligand;sFasL;ADAM10-processed FasL form;APL;FasL intracellular domain;FasL ICD;SPPL2A-processed FasL form;SPA;FASLG;APT1LG1, CD95L, FASL, TNFSF6;
Subcellular Localization Cell membrane ; Single-pass type II membrane protein . Cytoplasmic vesicle lumen . Lysosome lumen . Is internalized into multivesicular bodies of secretory lysosomes after phosphorylation by FGR and monoubiquitination (PubMed:17164290). Colocalizes with the SPPL2A protease at the cell membrane (PubMed:17557115). .
Molecular Weight 31485 MW

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein Function Cytokine that binds to TNFRSF6/FAS, a receptor that transduces the apoptotic signal into cells (PubMed:26334989, PubMed:9228058). Involved in cytotoxic T-cell-mediated apoptosis, natural killer cell-mediated apoptosis and in T-cell development (PubMed:9228058, PubMed:7528780, PubMed:9427603). Initiates fratricidal/suicidal activation-induced cell death (AICD) in antigen-activated T-cells contributing to the termination of immune responses (By similarity). TNFRSF6/FAS-mediated apoptosis has also a role in the induction of peripheral tolerance (By similarity). Binds to TNFRSF6B/DcR3, a decoy receptor that blocks apoptosis (PubMed:27806260). .
Background Fas ligand(FasL or CD95L) is a type-II transmembrane protein that belongs to the tumor necrosis factor(TNF) family. Its binding with its receptor induces apoptosis. The human FASL gene consists of approximately 8 kb and is split into 4 exons. Fas ligand/receptor interactions play an important role in the regulation of the immune system and the progression of cancer. Fas ligand or FasL is a homotrimeric type II transmembrane protein. It signals through trimerization of FasR, which spans the membrane of the "target" cell. This trimerization usually leads to apoptosis, or cell death. Soluble Fas ligand is generated by cleaving membrane-bound FasL at a conserved cleavage site by the external matrix metalloproteinase MMP-7.

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