HA724288

Human IL-1 alpha Recombinant Rabbit Monoclonal Antibody [PSH22-27]

cat.: HA724288

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human
Applications:	ELISA(Det)
Clonality:	Monoclonal
Clone number:	PSH22-27

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 IL-1 alpha aa 113-271.
Subcellular location:	Nucleus, Cytoplasm, Secreted.
Recommended Dilutions: ELISA(Det)	Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH22-26] to Human IL-1 alpha antibody (Capture) (HA724287) or Rabbit monoclonal [PSH22-28] to Human IL-1 alpha antibody (Capture) (HA724289) and Recombinant Human IL-1 alpha protein (HA210602) as the standard. The reference range value is 7.8-2,000 pg/mL.
Uniprot #:	SwissProt: P01583 Human
Alternative names:	BAF FAF Hematopoietin 1 Hematopoietin-1 IL 1 alpha IL 1A IL-1 alpha Il-1a IL1 ALPHA IL1 IL1A IL1A_HUMAN IL1F1 Interleukin 1 alpha Interleukin-1 alpha Interleukin1 alpha LAF LEM Preinterleukin 1 alpha Pro interleukin 1 alpha

Images

Fig1: Sandwich ELISA analysis of Human IL-1 alpha matched pair antibodies

Capture: HA724287, Human IL-1 alpha Rabbit mAb [PSH22-26]
Detector: HA724288, Human IL-1 alpha Rabbit mAb [PSH22-27]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724287) 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 IL-1 alpha protein (HA210602) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA724288, 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 IL-1 alpha matched pair antibodies

Capture: HA724289, Human IL-1 alpha Rabbit mAb [PSH22-28]
Detector: HA724288, Human IL-1 alpha Rabbit mAb [PSH22-27]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724289) 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 IL-1 alpha protein (HA210602) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA724288, 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 IL-1a in THP-1 cell culture supernatant untreated or treated with PMA and LPS for 24h. Capture: HA724287, Human IL-1 alpha Rabbit mAb [PSH22-26] Detector: HA724288, Human IL-1 alpha Rabbit mAb [PSH22-27] Interpolated concentration of native IL-1a was measured in duplicate at different sample concentrations and interpolated from the IL-1a standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean IL-1a concentration was determined to be 246 pg/ml in THP-1 treated cell culture supernatant and undetectable in THP-1 untreated cell culture supernatant.
	Fig4: Interpolated concentrations of native IL-1a in THP-1 cell culture supernatant untreated or treated with PMA and LPS for 24h. Capture: HA724289, Human IL-1 alpha Rabbit mAb [PSH22-28] Detector: HA724288, Human IL-1 alpha Rabbit mAb [PSH22-27] Interpolated concentration of native IL-1a was measured in duplicate at different sample concentrations and interpolated from the IL-1a standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean IL-1a concentration was determined to be 231 pg/ml in THP-1 treated cell culture supernatant and undetectable in THP-1 untreated cell culture supernatant.
	Fig5: Interpolated concentrations of native IL-1a in THP-1 cell extract samples based on a 1000 µg/ml extract load. Capture: HA724287, Human IL-1 alpha Rabbit mAb [PSH22-26] Detector: HA724288, Human IL-1 alpha Rabbit mAb [PSH22-27] Interpolated concentration of native IL-1a was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean IL-1a concentration was determined to be 7,724 pg/mL in THP-1 treated cell extract and undetectable in THP-1 untreated cell extract.

	Fig6: Interpolated concentrations of native IL-1a in THP-1 cell extract samples based on a 1000 µg/ml extract load. Capture: HA724289, Human IL-1 alpha Rabbit mAb [PSH22-28] Detector: HA724288, Human IL-1 alpha Rabbit mAb [PSH22-27] Interpolated concentration of native IL-1a was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean IL-1a concentration was determined to be 8,360 pg/mL in THP-1 treated cell extract and undetectable in THP-1 untreated cell extract.
	Fig7: Interpolated concentrations of spiked IL-1a in cell culture media samples. Capture: HA724287, Human IL-1 alpha Rabbit mAb [PSH22-26] Detector: HA724288, Human IL-1 alpha Rabbit mAb [PSH22-27] The concentrations of IL-1a were measured in duplicates, interpolated from the IL-1a 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 IL-1a in cell culture media samples. Capture: HA724289, Human IL-1 alpha Rabbit mAb [PSH22-28] Detector: HA724288, Human IL-1 alpha Rabbit mAb [PSH22-27] The concentrations of IL-1a were measured in duplicates, interpolated from the IL-1a 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”.