HA724023

Human Fas Ligand Recombinant Rabbit Monoclonal Antibody [PSH18-86]

cat.: HA724023

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human
Applications:	ELISA(Cap)
Clonality:	Monoclonal
Clone number:	PSH18-86

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4).
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Isotype:	IgG
Immunogen:	Recombinant protein within Human Fas Ligand aa 130-281.
Positive control:	Recombinant Human Fas Ligand protein.
Subcellular location:	Cell membrane, Cytoplasmic vesicle lumen, Lysosome lumen; Secreted; Nucleus.
Recommended Dilutions: ELISA(Cap)	Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH18-87] to Human Fas Ligand antibody (Detector) (HA724024) or Rabbit monoclonal [PSH18-88] to Human Fas Ligand antibody (Detector) (HA724025) and Recombinant Human Fas Ligand protein as the standard. The reference range value is 3.9-1,000 pg/mL.
Uniprot #:	SwissProt: P48023 Human
Alternative names:	ALPS1B Apoptosis (APO 1) antigen ligand 1 Apoptosis antigen ligand 1 Apoptosis antigen ligand APT1LG1 APTL CD178 CD178 antigen CD95 ligand CD95-L CD95L CD95L protein Fas antigen ligand Fas L Fas ligand (TNF superfamily member 6) Fas ligand FASL Fasl Fas ligand (TNF superfamily member 6) FASLG Generalized lymphoproliferative disease Gld soluble form TNFL6_HUMAN TNFSF6 Tumor necrosis factor (ligand) superfamily member 6 Tumor necrosis factor ligand superfamily member 6

Images

Fig1: Sandwich ELISA analysis of Human FASLG matched pair antibodies

Capture: HA724023, Human Fas Ligand Rabbit mAb [PSH18-86]
Detector: HA724024, Human Fas Ligand Rabbit mAb [PSH18-87]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724023) diluted in carbonate/bicarbonate buffer, at a concentration of 5 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant Human Fas Ligand protein starting from 1,000 pg/ml to 0 pg/ml and detect antibody (HA724024, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

Fig2: Sandwich ELISA analysis of Human FASLG matched pair antibodies

Capture: HA724023, Human Fas Ligand Rabbit mAb [PSH18-86]
Detector: HA724025, Human Fas Ligand Rabbit mAb [PSH18-88]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724023) diluted in carbonate/bicarbonate buffer, at a concentration of 5 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant Human Fas Ligand protein starting from 1,000 pg/ml to 0 pg/ml and detect antibody (HA724025, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

	Fig3: Interpolated concentrations of native FASLG in NK-92 cell culture supernatant. Capture: HA724023, Human Fas Ligand Rabbit mAb [PSH18-86] Detector: HA724024, Human Fas Ligand Rabbit mAb [PSH18-87] Interpolated concentration of native FASLG was measured in duplicate at different sample concentrations and interpolated from the FASLG standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean FASLG concentration was determined to be 1,077 pg/ml in NK-92 cell culture supernatant.
	Fig4: Interpolated concentrations of native FASLG in NK-92 cell culture supernatant. Capture: HA724023, Human Fas Ligand Rabbit mAb [PSH18-86] Detector: HA724025, Human Fas Ligand Rabbit mAb [PSH18-88] Interpolated concentration of native FASLG was measured in duplicate at different sample concentrations and interpolated from the FASLG standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean FASLG concentration was determined to be 890 pg/ml in NK-92 cell culture supernatant.
	Fig5: Interpolated concentrations of native FASLG in NK-92 Cell extract samples based on a 1000 µg/ml extract load. Capture: HA724023, Human Fas Ligand Rabbit mAb [PSH18-86] Detector: HA724024, Human Fas Ligand Rabbit mAb [PSH18-87] Interpolated concentration of native FASLG was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean FASLG concentration was determined to be 1,191 pg/ml in NK-92 treated Cell extract and 493 pg/ml in NK-92 untreated Cell extract.

	Fig6: Interpolated concentrations of native FASLG in NK-92 Cell extract samples based on a 1000 µg/ml extract load. Capture: HA724023, Human Fas Ligand Rabbit mAb [PSH18-86] Detector: HA724025, Human Fas Ligand Rabbit mAb [PSH18-88] Interpolated concentration of native FASLG was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean FASLG concentration was determined to be 2,182 pg/ml in NK-92 treated Cell extract and 540 pg/ml in NK-92 untreated Cell extract.
	Fig7: Interpolated concentrations of spiked FASLG in cell culture media samples. Capture: HA724023, Human Fas Ligand Rabbit mAb [PSH18-86] Detector: HA724024, Human Fas Ligand Rabbit mAb [PSH18-87] The concentrations of FASLG were measured in duplicates, interpolated from the FASLG standard curves and corrected for sample dilution. diluted samples are as follows: 50% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
	Fig8: Interpolated concentrations of spiked FASLG in cell culture media samples. Capture: HA724023, Human Fas Ligand Rabbit mAb [PSH18-86] Detector: HA724025, Human Fas Ligand Rabbit mAb [PSH18-88] The concentrations of FASLG were measured in duplicates, interpolated from the FASLG standard curves and corrected for sample dilution. diluted samples are as follows: 50% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.