16-9919-85

antibody from Invitrogen Antibodies

Targeting: FASLG APT1LG1, CD178, FasL, TNFSF6

Flow cytometry (Supportive data in Antibodypedia)

Blocking/Neutralizing (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

16-9919-85

Provider

Invitrogen Antibodies

Product name

CD178 (Fas Ligand) Monoclonal Antibody (NOK-1), Functional Grade, eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The NOK-1 monoclonal antibody reacts with human Fas (CD95) Ligand, a 40 kDa type II transmembrane glycoprotein. FasL is a member of the TNF family and is expressed by neutrophils, monocytes, and activated T cells and NK cells. The interaction of FasL with its receptor (CD95, Fas) induces Fas-mediated killing of lymphocytes. Human FasL is cleaved from the surface by matrix metalloproteinases (MMPs), resulting in a 26 kDa soluble form. Therefore for optimal detection of surface FasL on activated peripheral blood cells, incubation of cells with an MMP inhibitor is recommended. Applications Reported: NOK-1 has been reported for use in flow cytometric analysis. NOK-1 has also been reported in blocking of FasL mediated killing in functional assays. Applications Tested: The NOK-1 antibody has been tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at less than or equal to 1 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Storage and handling: Use in a sterile environment. Filtration: 0.2 µm post-manufacturing filtered. Purity: Greater than 90%, as determined by SDS-PAGE. Endotoxin Level: Less than 0.001 ng/µg antibody, as determined by LAL assay. Aggregation: Less than 10%, as determined by HPLC.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

NOK-1

Vial size

500 µg

Concentration

1 mg/mL

Storage

4° C

References

Submitted references

CHMP2A regulates tumor sensitivity to natural killer cell-mediated cytotoxicity.

Bernareggi D, Xie Q, Prager BC, Yun J, Cruz LS, Pham TV, Kim W, Lee X, Coffey M, Zalfa C, Azmoon P, Zhu H, Tamayo P, Rich JN, Kaufman DS
Nature communications 2022 Apr 7;13(1):1899

---

CD19-targeted CAR regulatory T cells suppress B cell pathology without GvHD.

Imura Y, Ando M, Kondo T, Ito M, Yoshimura A
JCI insight 2020 Jul 23;5(14)

---

Transmembrane TNF-α promotes activation-induced cell death by forward and reverse signaling.

Zhang M, Wang J, Jia L, Huang J, He C, Hu F, Yuan L, Wang G, Yu M, Li Z
Oncotarget 2017 Sep 8;8(38):63799-63812

---

Functional dichotomy of Vδ2 γδ T cells in chronic hepatitis C virus infections: role in cytotoxicity but not for IFN-γ production.

Yin W, Tong S, Zhang Q, Shao J, Liu Q, Peng H, Hu H, Peng M, Hu P, Ren H, Tian Z, Zhang D
Scientific reports 2016 May 19;6:26296

---

The probiotic Escherichia coli strain Nissle 1917 induces gammadelta T cell apoptosis via caspase- and FasL-dependent pathways.

Guzy C, Paclik D, Schirbel A, Sonnenborn U, Wiedenmann B, Sturm A
International immunology 2008 Jul;20(7):829-40

---

Membrane Fas ligand kills human peripheral blood T lymphocytes, and soluble Fas ligand blocks the killing.

Suda T, Hashimoto H, Tanaka M, Ochi T, Nagata S
The Journal of experimental medicine 1997 Dec 15;186(12):2045-50

---

Expression of the functional soluble form of human fas ligand in activated lymphocytes.

Tanaka M, Suda T, Takahashi T, Nagata S
The EMBO journal 1995 Mar 15;14(6):1129-35

---

Metalloproteinase-mediated release of human Fas ligand.

Kayagaki N, Kawasaki A, Ebata T, Ohmoto H, Ikeda S, Inoue S, Yoshino K, Okumura K, Yagita H
The Journal of experimental medicine 1995 Dec 1;182(6):1777-83

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Staining of human FasL tranfected cells with Anti-Human CD178 (CD95 Ligand) PE. Appropriate isotype controls were used (open histogram). Total viable cells were used for analysis.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

EVs secreted by Cal27 induce apoptosis in NK cells. a Cal27-WT and KO derived EVs were analyzed by flow cytometry for known NKG2D ligands MICA/B, ULBP2/5/6, ULPB3, and for TRAIL and FasL, proteins involved in intrinsic or extrinsic apoptosis pathway (green histograms). In blue is shown the isotype control. Representative of n = 2 independent experiments. b Flow cytometry CASP3/7 viability assay showing increasing NK cell death once exposed to EVs from Cal27 in a time course of 4-hours. The error bars represent +-SEM across n = 3 replicates. Two-way ANOVA and Bonferroni's post hoc test was performed to determine statistical significance. Representative of n = 3 independent experiments. c 24 hour time course viability assay quantified using the IncuCyte real-time imaging system. N = 5 images/well taken for each of three replicates. The error bars represent +-SEM across n = 3 replicates. Statistical analysis was performed by two-way ANOVA and Bonferroni's post hoc multiple comparison test. Representative of n = 2 independent experiments. d 4 hour cytotoxicity assay using NK cells as effectors against Cal27-WT or KO as target cells. Here we show how Cal27-derived EVs reduce the killing in Cal27-KO. Error bars represent +-SEM across n = 3 replicates. Statistical analysis was performed by one-way ANOVA and Bonferroni's post hoc test (ns, not significant). e antibodies blocking MICA/B, TRAIL, or a combination of the two (aMICA/B and aTRAIL) limit the inhibitory effect of EV on NK ce