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| SKU | EK0338 |
|---|---|
| Size | 96wells/kit, with removable strips. |
| Reactivity | Mouse |
| Sample Type | cell culture supernates, serum and plasma(heparin, EDTA, citrate). |
| Sensitivity | <10pg/ml |
| Assay Range | 31.2pg/ml-2000pg/ml |
Overview
| Product Name | Mouse FASL PicoKine™ ELISA Kit |
|---|---|
| SKU/Catalog Number | EK0338 |
| 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 | Sandwich High Sensitivity ELISA kit for Quantitative Detection of Mouse FASL. 96wells/kit, with removable strips. |
| Cite This Product | Mouse FASL PicoKine™ ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0338) |
| 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 thata are not specified above or contact us to check for feasibility. |
| Sensitivity | <10pg/ml |
| Assay Range | 31.2pg/ml-2000pg/ml |
| Immunogen | Expression system for standard: NSO; Immunogen sequence: P132-L279 |
| Reactivity | Mouse |
| Cross Reactivity | There is no detectable cross-reactivity with other relevant proteins. |
Assay Details
Kit Components
| Catalog number | Description | Quantity |
| EK0338-CAP | 96-well plate precoated with anti-Mouse Faslg antibody | 1 |
| EK0338-ST | lyophilized recombinant Mouse Faslg standard | 10ng/tube |
| EK0338-DA | biotinylated anti-Mouse Faslg antibody | 130ul(dilution 1:100) |
| AR1103 | Avidin-Biotin-Peroxidase Complex(ABC) | 130ul(dilution 1:100) |
| AR1106-1 | sample diluent buffer | 30ml |
| AR1106-2 | antibody diluent buffer | 12ml |
| AR1106-3 | ABC diluent buffer | 12ml |
| AR1104 | TMB color developing agent | 10ml |
| AR1105 | TMB stop solution | 10ml |
| PLA-SEA | Adhesive cover | 4 |
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 Mouse FASL PicoKine™ ELISA Kit
(TMB reaction incubate at 37°C for 20-25min)
| Concentration(pg/ml) | 0 | 2000 | 2000 | 2000 | 2000 | 2000 | 2000 | 2000 |
| O.D. | 0.016 | 0.074 | 0.127 | 0.232 | 0.456 | 0.863 | 1.521 | 2.210 |
Intra/Inter Assay Precision
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean(pg/ml) | 68 | 206 | 772 | 65 | 216 | 727 |
| Standard deviation | 2.99 | 13.59 | 49.4 | 3.51 | 17.28 | 47.98 |
| CV(%) | 4.4% | 6.6% | 6.4% | 5.4% | 8% | 6.6% |
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.
| Lots | Lot1 (pg/ml) | Lot2 (pg/ml) | Lot3 (pg/ml) | Lot4 (pg/ml) | Mean (pg/ml) | Standard Deviation | CV (%) |
|---|---|---|---|---|---|---|---|
| Sample 1 | 68 | 63 | 68 | 59 | 64 | 3.77 | 5.8% |
| Sample 2 | 206 | 196 | 205 | 225 | 208 | 10.55 | 5% |
| Sample 3 | 772 | 915 | 764 | 889 | 835 | 67.68 | 8.1% |
*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 | P41047 |
|---|---|
| Gene Name | Faslg |
| Protein Name | Tumor necrosis factor ligand superfamily member 6 |
| Tissue Specificity | Expressed in T-cells (PubMed:19794494). Expressed in natural killer cells (PubMed:7532682). . |
| Alternative Names | Tumor necrosis factor ligand superfamily member 6;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, gld, Tnfsf6; |
| Subcellular Localization | Isoform FasL: 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. Colocalizes with the SPPL2A protease at the cell membrane. . |
| Molecular Weight | 31442 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:7511063). Involved in cytotoxic T-cell-mediated apoptosis, natural killer cell-mediated apoptosis and in T-cell development (PubMed:19794494, PubMed:7532682). Initiates fratricidal/suicidal activation-induced cell death (AICD) in antigen-activated T-cells contributing to the termination of immune responses (PubMed:19794494). TNFRSF6/FAS-mediated apoptosis has also a role in the induction of peripheral tolerance (PubMed:10779162). Binds to TNFRSF6B/DcR3, a decoy receptor that blocks apoptosis (By similarity). . |
|---|---|
| Background | FAS Ligand(FASL) is a 40 kDa type II membrane protein belonging to the tumor necrosis factor family, which induces apoptosis by binding to its receptor, Fas. The human FasL gene consists of approximately 8.0 kb and is split into four exons. This gene consists of 281 amino acids with a calculated M(r) of 31,759 and was mapped on chromosome 1q23. It has an identity of 76.9% at the amino acid sequence level with mouse FasL. The FAS and FASL system plays a key role in regulating apoptotic cell death and corruption of this signalling pathway has been shown to participate in immune escape and tumorigenesis. FAS and FASL triggered apoptosis pathway plays an important role in human carcinogenesis. This system may also play a role in modulating the genetic susceptibility of mouse strains to develop T-cell lymphoblastic lymphomas. |
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Publications
Cisplatin-induced oxidative stress stimulates renal Fas ligand shedding
The upregulation of integrin ?D?2 (CD11d/CD18) on inflammatory macrophages promotes macrophage retention in vascular lesions and development of atherosclerosis
Han X, Qiu L, Zhang Y, Kong Q, Wang H, Wang H, Li H, Duan C, Wang Y, Song Y, Wang C. Exp Biol Med (Maywood). 2009 May;234(5):595-603. Doi: 10.3181/0812-Rm-359. Epub 2009 Feb 25. Transplantation Of Sertoli-Islet Cell Aggregates Formed By Microgravi...
Xu J, Liu H, Wu Y, Gong X, Zhou Q, Qiao F. J Obstet Gynaecol Res. 2011 Mar;37(3):187-94. Doi: 10.1111/J.1447-0756.2010.01334.X. Epub 2010 Dec 16. Proapoptotic Effect Of Metalloproteinase 9 Secreted By Trophoblasts On Endothelial Cells.
Xiong M, Lu J, Zhao A, Xu L, Bao S, Lin Q, Yang B. Fertil Steril. 2010 May 15;93(8):2767-9. Doi: 10.1016/J.Fertnstert.2009.11.040. Epub 2010 Feb 1. Therapy With Fasl-Gene-Modified Dendritic Cells Confers A Protective Microenvironment In Murine Pre...
Wu Yh, Hu Sq, Liu J, Cao Hc, Xu W, Li Yj, Li Lj. Int J Mol Med. 2014 Jun;33(6):1498-506. Doi: 10.3892/Ijmm.2014.1730. Epub 2014 Apr 7. Nature And Mechanisms Of Hepatocyte Apoptosis Induced By D-Galactosamine/Lipopolysaccharide Challenge In Mice.
Customer Q&As
Q: Is it possible to dilute 10X TBS stock that is 250mM Tris Base/Tris-Hcl, 27 mM KCl, and 1.37M NaCl to be used as the wash buffer?
A: You can dilute the 10X TBS stock to 1XTBS and test the pH value of the 1xTBS.
The 1XTBS can be used if the pH value falls in the range of 7.2-7.6.
The 1XTBS can be used if the pH value falls in the range of 7.2-7.6.
Q: Would your kits work normall with samples from Wistar Rat?
A: Our kits would work normally with samples from Wistar Rat.
Q: How can I store the dissolved wash buffer for a longer period of time?
A: You can prepare a concentrated wash buffer (25X) which can be stored at 4°C for 1-3 months.
Q: Does the TMB color developing agent contain H2O2?
A: Yes, the TMB color developing agent contain H2O2.
Q: Does this ELISA kit contain any product produced in humans or primates?
Keywords: component, ingredient
Keywords: component, ingredient
A: None of the components in our ELISA kits are produced in humans or primates.
Q: What is the volume of the recombinant protein control? What is its shelf life?
Keywords: expiration, storage, temperature, size
Keywords: expiration, storage, temperature, size
A: The volume of the control is around 100pg. If unopened, the shelf life of the control is the same as the whole kit - "Store at 4˚C for 6 months, at -20˚C for 12 months." The reconstituted control can be stored at -20˚C for 2 days.
Q: Will it be okay to run the experiment if I accidentally used the wrong buffer (e.g. sample diluent buffer) for antibody dilution instead of the antibody diluent buffer? Keywords: dilution, replace, substitute, sample buffer
A: Using the sample diluent buffer for antibody dilution instead of the antibody diluent buffer will negatively affect the test result to some extent. The values of both standard and sample might be lower than the normal values.
Q: The protocol says to use neutral PBS and provides a recipe. Could I use neutral PBS made with KCl or KH2PO4 (common constituents for most PBS recipes) instead?
Keywords: protocol, alternative buffer, applications, reagent recipe
A: Yes, it is ok to use common PBS recipes. We have tried many types of buffers with common constituents, and no significant difference was observed whatsoever.
Q: Why are my O.D. values different than your values on the datasheet?
Keyword: troubleshooting, reference
A: We detected the kit in our lab and got our values on the datasheet before delivery, but protein activity will decrease as time goes on, so you may get lower O.D. values than ours. However, it should still be in a reasonable range and the standards can be used to calculate sample values. Good linearity of curve is more important than the actual numerical value.
Q: On the ELISA kit datasheet it says "HRP substrate TMB was used to visualize HRP enzymatic reaction." What is HRP and which part of the kit contains HRP?
Keyword: kit components, reagents, protocol
A: It means Avidin-Biotin-Peroxidase Complex(AR1103) and you could find it in Kit Components.
Q: Do you offer any ELISA kits that can work with whole blood samples instead of plasma or urine?
Keyword: applications
A: We test serum and plasma routinely, and there is very little difference between serum, plasma and whole blood. The whole blood also contains proteins we need to test. The kit can be used to test whole blood in theory.
Q: Does silicic acid (formula SiOH4) affect the results of the ELISA assay?
Keyword: protocol, reagents
A: The acid may affect the binding of antigen to antibody, it is not recommended to use. That being said, it is unlikely to affect the reaction if the solution remains an overall neutral environment.
Q: Is there lot-to-lot variation of the ELISA kits? What are Boster's general services when I have questions about your kits?
Keyword: technical support, help, customer service
A: The NIBSC/WHO 1st International Standard is evaluated, we always test our kits before delivery, and customers can find the test result on the protocol. If the customer needs technical support from us to analyze their data, please contact us and we ask that you provide us the following information: the lot# and production date, and when did the customer detect the kit.
Q: The results of my standards do not look correct, what could be the problem?
Keyword: troubleshoot, protocol
A: Double check the protocol for plate washing. It should be examined on three aspects: 1) Did you make the washing buffer correctly? 2) How long did you let the buffer stay in wells? 3) What was the volume of the washing buffer added to each well? In addition, pay attention when adding sample to avoid contamination. And we suggest testing the standards again. Values of standards at low concentration are more affected than that at higher concentration, so customer can still get expected values at high concentration even if there is an error. If problems persist, please contact technical support with the catalog and lot number of your product.
Q: What is the well depth of the 96 well plates for the ELISA kits?
Keyword: well capacity, product size
A: The well depth is 300ul, and the max capacity is 350ul. The height of the well is 1.1 cm, and the internal size is 1 cm.
Q: The kit does not include wash solution, what should I do?
Keyword: wash buffer
A: Our Elisa kits do not come with wash solution, but we have included information about how to make washing buffer on the datasheet. Please refer to the "Material Required But Not Provided" section. We also offer washing buffer for sale separately (Phosphate Buffered Saline Powder SKU: AR0030).
Q: For your ELISA kits, are the capture and detection antibodies polyclonals or monoclonals?
Keyword: antibody clonality
A: This information can be found for each kit under the "Properties" section, and you can find the immunogen sequence information in the "Overview" section.
Q: Do your ELISA kits come with sealants or plate covers?
Keyword: storage, sequential use
A: The kit can be used within a month sequentially if it's opened and stored at 4 degree. There is no need to use sealants, the plate can be packaged with aluminum foil bag, and for other reagents keep bottle tightly closed.
Q: What is the concentration of Sodium Azide in your Elisa kits?
Keyword: preservative
A: Our Elisa kits contain 0.02% Sodium Azide. All of the components except TMB colour developing reagent and stop solution contains 0.02% Sodium azide as the preservative.
Q: Are there positive and negative controls available for my ELISA kit?
Keyword: positive control, negative control
A: We can provide a recombinant protein as a control. It costs $50 per control and takes 2 weeks to manufacture. We cannot provide a positive or negative control in serum.
Q: Why do I get positive results for my knockout (KO) model when used as a control?
A: The knockout (KO) model may contain truncated forms of the target protein which can be detected by the ELISA assay.
Q: How long do I soak my plate in the wash buffer?
A: The plate should soak in 0.3mL PBS or TBS wash buffer for at least 1-2 minutes in an automated wash station.
Q: Can I use Tween in my wash buffer?
A: While it is not recommended to use Tween in your wash buffer, small amounts (<0.1% concentration) may decrease background due to insufficient blocking.
Q: What plate type do I use to set up the microplate reader?
A: Our plates are made with the Corning costar plates similar to the DNA-BIND 96 -well plates.
Q: What should I use for negative control?
A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the normal level of this protein in my sample of interest?
A: We have reviewed literature and have gathered this information for most of our ELISA kits. You can find this information on the product page or contact us if you cannot find it. However this information is only suggestive and cannot replace pilot studies in determining the optimal sample dilution ratio.
Q: Is the plate separable? Can I use only part of the kit and save the rest for later? How many samples can I run with one kit?
A: Yes the plate is separable. There are 12 strips of 8 wells each, all removable from the plate. The amount of samples you can run depend on a few factors. In the most common ELISA set up, you will use two strips for standards, and 10 strips for samples using duplicates, which let you run up to 40 samples per kit. Contact us if you have questions regarding other situations.
Q: Does the kit contain sample preparation reagents? How do I prepare my samples?
A: Since different sample types require different reagents, we do not include them in the ELISA kit. However we do have each reagent mentioned in the file below available at very reasonable prices. Be sure to check them out. For sample preparation protocols please download the file below: https://www.bosterbio.com/media/pdf/Boster_Sample_Preparation_Protocols.pdf
Q: Can this ELISA kit work on brain tissue homogenate, cell culture supernatant, saliva, urine, serum, whole blood or any other sample type?
A: In theory the ELISA kit will work for all sample types if the target protein is present at a level that falls within the linear range of the ELISA kit detection range. We guarantee the kit to work on the sample types that we have tested. If you want dilution ratio suggestions on these sample types please contact our technical support. For sample types that we have not tested for, we suggest you run pilot experiments to decide the optimal sample dilution.
Q: Can this ELISA kit react with the pro-form of the target protein? Can this ELISA kit react with an isoform of the protein?
A: In general, unless otherwise specified, the ELISA kit is pan-specific, meaning that it will react with all different forms of the target protein if they share the majority of the target protein's sequence. The capture and detection antibodies are reactive to the entire sequence of the standard protein. You can find the sequence information of the standard protein in the "immunogen" section. For more information about the specificity of the kit for your particular experiment, please contact our techincal support.
Q: Can this ELISA kit react with human, mouse, rat or other species?
A: If the kit is reactive to another commonly used species (human, mouse, and/or rat), we would have listed it as a separate product. If you want to check cross-reactivity to a species that is not included in the 3 species listed above, please contact our technical support for more information. As a rule of thumb, if the sequences are 90%+ identical, there is a high chance of cross-reactivity for your species of interest.
Q: What are some alternative names that could be used to describe this product?
A: One other very common name is fas elisa kit