Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH14-58] to Human Endoglin / CD105 antibody (Capture) (HA723654) or Rabbit monoclonal [PSH14-60] to Human Endoglin / CD105 antibody (Capture) (HA723657) and Recombinant Human Endoglin / CD105 protein (HA211006) as the standard. The reference range value is 19.5-5,000 pg/mL.
ELISA
Use at an assay dependent concentration.
Target
Function
Endoglin (ENG) is a type I membrane glycoprotein located on cell surfaces and is part of the TGF beta receptor complex. It is also commonly referred to as CD105, END, FLJ41744, HHT1, ORW and ORW1. It has a crucial role in angiogenesis, therefore, making it an important protein for tumor growth, survival and metastasis of cancer cells to other locations in the body. Endoglin has been found to be an auxiliary receptor for the TGF-beta receptor complex. It thus is involved in modulating a response to the binding of TGF-beta1, TGF-beta3, activin-A, BMP-2, BMP-7 and BMP-9. Beside TGF-beta signaling endoglin may have other functions. It has been postulated that endoglin is involved in the cytoskeletal organization affecting cell morphology and migration. Endoglin has a role in the development of the cardiovascular system and in vascular remodeling. Its expression is regulated during heart development . Experimental mice without the endoglin gene die due to cardiovascular abnormalities.
Background References
1. Litwiniuk-Kosmala M et al. Endoglin in head and neck neoplasms. Front Med (Lausanne). 2023 Feb
2. González Muñoz T et al. Endoglin in the Spotlight to Treat Cancer. Int J Mol Sci. 2021 Mar
Sandwich ELISA analysis of Human Endoglin / CD105 matched pair antibodies
Capture: HA723654, Human Endoglin / CD105 Rabbit mAb [PSH14-58] Detector: HA723655, Human Endoglin / CD105 Rabbit mAb [PSH14-59]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723654) diluted in carbonate/bicarbonate buffer, at a concentration of 2 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 Endoglin / CD105 protein (HA211006) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA723655, 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.
Sandwich ELISA analysis of Human Endoglin / CD105 matched pair antibodies
Capture: HA723657, Human Endoglin / CD105 Rabbit mAb [PSH14-60] Detector: HA723655, Human Endoglin / CD105 Rabbit mAb [PSH14-59]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723657) diluted in carbonate/bicarbonate buffer, at a concentration of 2 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 Endoglin / CD105 protein (HA211006) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA723655, 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.
Interpolated concentrations of native Endoglin / CD105 in Human serum and Jurkat supernatant.
Capture: HA723654, Human Endoglin / CD105 Rabbit mAb [PSH14-58] Detector: HA723655, Human Endoglin / CD105 Rabbit mAb [PSH14-59]
Interpolated concentration of native Endoglin / CD105 was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Endoglin / CD105 concentration was determined to be 3975 pg/ml in Human serum,undetectable in Jurkat cell supernatant.
Interpolated concentrations of native Endoglin / CD105 in Human serum and Jurkat cell culture supernatant.
Capture: HA723657, Human Endoglin / CD105 Rabbit mAb [PSH14-60] Detector: HA723655, Human Endoglin / CD105 Rabbit mAb [PSH14-59]
Interpolated concentration of native Endoglin / CD105 was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Endoglin / CD105 concentration was determined to be 3082pg/ml in Human serum,undetectable in Jurkat cell culture supernatant.
Interpolated concentrations of native Endoglin / CD105 in HUVEC, Jurkat and Raji Cell extract samples based on a 1000 µg/ml extract load.
Capture: HA723654, Human Endoglin / CD105 Rabbit mAb [PSH14-58] Detector: HA723655, Human Endoglin / CD105 Rabbit mAb [PSH14-59]
Interpolated concentration of native Endoglin / CD105 was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Endoglin / CD105 concentration was determined to be 3207 pg/ml in HUVEC Cell extract,undetectable in Jurkat and Raji Cell extract.
Interpolated concentrations of native Endoglin / CD105 in HUVEC, Jurkat and Raji Cell extract samples based on a 1000 µg/ml extract load.
Capture: HA723657, Human Endoglin / CD105 Rabbit mAb [PSH14-60] Detector: HA723655, Human Endoglin / CD105 Rabbit mAb [PSH14-59]
Interpolated concentration of native Endoglin / CD105 was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Endoglin / CD105 concentration was determined to be 3462 pg/ml in HUVEC Cell extract,undetectable in Jurkat and Raji Cell extract.
Interpolated concentrations of spiked ENG in cell culture media samples.
Capture: HA723654, Human Endoglin / CD105 Rabbit mAb [PSH14-58] Detector: HA723655, Human Endoglin / CD105 Rabbit mAb [PSH14-59]
The concentrations of Endoglin / CD105 were measured in duplicates, interpolated from the Endoglin / CD105 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
Interpolated concentrations of spiked Endoglin / CD105 in cell culture media samples.
Capture: HA723657, Human Endoglin / CD105 Rabbit mAb [PSH14-60] Detector: HA723655, Human Endoglin / CD105 Rabbit mAb [PSH14-59]
The concentrations of Endoglin / CD105 were measured in duplicates, interpolated from the Endoglin / CD105 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"