Biotin Conjugated Human Caspase-1 Recombinant Rabbit Monoclonal Antibody [PSH24-90]
cat.: HA724485B
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: ELISA(Det), ELISA
Clonality: Monoclonal
Clone number: PSH24-90
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage buffer: 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.05% Proclean 300.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Isotype: IgG
Immunogen: Recombinant protein within Human Caspase-1 aa 120-404 (HA211426).
Positive control: Recombinant Human Caspase-1 protein (HA211426).
Subcellular location: Cytoplasm, Cell membrane.
Recommended Dilutions:
  ELISA(Det)
  ELISA
Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH24-89] to Human Caspase-1 (Capture) (HA724484) or Rabbit monoclonal [PSH24-91] to Human Caspase-1 (Capture) (HA724486) and Recombinant Human Caspase-1 protein (HA211426) as the standard. The reference range value is 23.4-3,000 pg/mL.
Use at an assay dependent concentration.
Uniprot #: SwissProt: P29466 Human
Alternative names: Caspase1 CASP-1 CASP1 CASP1_HUMAN Caspase1 Caspase 1 Caspase-1 subunit p10 ICE IL-1 beta-converting enzyme IL-1BC IL1 beta converting enzyme IL1B convertase Interleukin 1 beta convertase Interleukin 1B converting enzyme Interleukin-1 beta convertase Interleukin-1 beta-converting enzyme p45
Images
HA724485B_1.png Fig1: Sandwich ELISA analysis of Human Caspase-1 matched pair antibodies

Capture: HA724484, Human Caspase-1 Rabbit mAb [PSH24-89]
Detector: HA724485, Human Caspase-1 Rabbit mAb [PSH24-90]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724484) 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 Caspase-1 protein (HA211426) starting from 3,000 pg/mL to 0 pg/mL and detect antibody (HA724485, 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.
HA724485B_2.png Fig2: Sandwich ELISA analysis of Human Caspase-1 matched pair antibodies

Capture: HA724486, Human Caspase-1 Rabbit mAb [PSH24-91]
Detector: HA724485, Human Caspase-1 Rabbit mAb [PSH24-90]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724486) 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 Caspase-1 protein (HA211426) starting from 3,000 pg/mL to 0 pg/mL and detect antibody (HA724485, 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.
HA724485B_3.png Fig3: Interpolated concentrations of native Caspase-1 in THP-1, HL-60 and HEK-293 cell extract samples based on a 1,000 µg/ml extract load.

Capture: HA724484, Human Caspase-1 Rabbit mAb [PSH24-89]
Detector: HA724485, Human Caspase-1 Rabbit mAb [PSH24-90]

Interpolated concentration of native Caspase-1 was measured in duplicate at different sample concentrations and interpolated from the Caspase-1 standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Caspase-1 concentration was determined to be 11,357 pg/mL in THP-1 cell extract and 4,280 pg/mL in HL-60 cell extract. There was no detectable signal in HEK-293 cell extract.
HA724485B_4.png Fig4: Interpolated concentrations of native Caspase-1 in THP-1, HL-60 and HEK-293 cell extract samples based on a 1,000 µg/ml extract load.

Capture: HA724486, Human Caspase-1 Rabbit mAb [PSH24-91]
Detector: HA724485, Human Caspase-1 Rabbit mAb [PSH24-90]

Interpolated concentration of native Caspase-1 was measured in duplicate at different sample concentrations and interpolated from the Caspase-1 standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Caspase-1 concentration was determined to be 16,889 pg/mL in THP-1 cell extract and 4,216 pg/mL in HL-60 cell extract. There was no detectable signal in HEK-293 cell extract.
HA724485B_5.png Fig5: Interpolated concentrations of spiked Caspase-1 in cell culture media samples.

Capture: HA724484, Human Caspase-1 Rabbit mAb [PSH24-89]
Detector: HA724485, Human Caspase-1 Rabbit mAb [PSH24-90]

The concentrations of Caspase-1 were measured in duplicates, interpolated from the Caspase-1 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).
HA724485B_6.png Fig6: Interpolated concentrations of spiked Caspase-1 in cell culture media samples.

Capture: HA724486, Human Caspase-1 Rabbit mAb [PSH24-91]
Detector: HA724485, Human Caspase-1 Rabbit mAb [PSH24-90]

The concentrations of Caspase-1 were measured in duplicates, interpolated from the Caspase-1 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”.