HA724128

Human/Monkey Albumin Recombinant Rabbit Monoclonal Antibody [PSH20-17]

cat.: HA724128

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Dog, Cynomolgus monkey
Applications:	ELISA(Det)
Clonality:	Monoclonal
Clone number:	PSH20-17

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 Serum Albumin.
Positive control:	Human Albumin protein (A3782, sigma).
Subcellular location:	Secreted.
Recommended Dilutions: ELISA(Det)	Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH20-16] to Human/Monkey Albumin antibody (Capture) (HA724127) and Human Albumin protein (A3782, sigma) as the standard. The reference range value is 78.1-5,000 pg/mL.
Uniprot #:	SwissProt: P02768 Human
Alternative names:	alb ALBU_HUMAN Albumin (32 AA) Albumin (AA 34) cell growth inhibiting protein 42 DKFZp779N1935 GIG20 GIG42 growth-inhibiting protein 20 OTTHUMP00000220436 OTTHUMP00000220438 OTTHUMP00000220439 PRO0883 PRO0903 PRO1341 PRO1708 PRO2044 PRO2619 PRO2675 Serum albumin UNQ696/PRO1341

Images

Fig1: Sandwich ELISA analysis of Human Albumin matched pair antibodies

Capture: HA724127, Human/Monkey Albumin Rabbit mAb [PSH20-16]
Detector: HA724128, Human/Monkey Albumin Rabbit mAb [PSH20-17]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724127) diluted in carbonate/bicarbonate buffer, at a concentHumanion 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 Human Albumin protein (A3782, sigma) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA724128, Biotin, 0.05 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 SubstHumane 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 Monkey Albumin matched pair antibodies

Capture: HA724127, Human/Monkey Albumin Rabbit mAb [PSH20-16]
Detector: HA724128, Human/Monkey Albumin Rabbit mAb [PSH20-17]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724127) diluted in carbonate/bicarbonate buffer, at a concentCynomolgusion 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 Monkey Albumin protein (BSA-CM101, Kactusbio) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA724128, Biotin, 0.05 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 SubstHumane 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 Albumin in human serum samples. Capture: HA724127, Human/Monkey Albumin Rabbit mAb [PSH20-16] Detector: HA724128, Human/Monkey Albumin Rabbit mAb [PSH20-17] The concentrations of Albumin were measured in duplicates, interpolated from the Albumin standard curve and corrected for sample dilution. Undiluted samples are human serum 0.0000063%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Albumin concentration was determined to be 25.6 mg/ml in human serum.
	Fig4: Interpolated concentrations of native Albumin in HepG2 and HeLa cell culture supernatant. Capture: HA724127, Human/Monkey Albumin Rabbit mAb [PSH20-16] Detector: HA724128, Human/Monkey Albumin Rabbit mAb [PSH20-17] The concentrations of Albumin were measured in duplicates, interpolated from the Albumin standard curve and corrected for sample dilution. Undiluted samples are HepG2 cell culture supernatant 0.5% and HeLa cell culture supernatant 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Albumin concentration was determined to be 745.1 ng/ml in HepG2 cell culture supernatant and undetectable in HeLa cell culture supernatant.
	Fig5: Interpolated concentrations of spiked Albumin in human cell culture media samples. Capture: HA724127, Human/Monkey Albumin Rabbit mAb [PSH20-16] Detector: HA724128, Human/Monkey Albumin Rabbit mAb [PSH20-17] The concentrations of Albumin were measured in duplicates, interpolated from the Albumin standard curves and corrected for sample dilution. Diluted samples are as follows: 25% 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”.