Biotin Conjugated Mouse ICAM1 Recombinant Rabbit Monoclonal Antibody [PSH16-35]
cat.: HA723833B
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Mouse
Applications: ELISA(Det), ELISA
Clonality: Monoclonal
Clone number: PSH16-35
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. 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 Mouse ICAM1 aa 24-485 (HA211066).
Positive control: Recombinant Mouse ICAM1 protein (HA211066).
Subcellular location: Membrane.
Recommended Dilutions:
  ELISA(Det)
  ELISA
Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH16-34] to Mouse ICAM1 antibody (Capture) (HA723832) or Rabbit monoclonal [PSH16-36] to Mouse ICAM1 antibody (Capture) (HA723834) and Recombinant Mouse ICAM1 protein (HA211066) as the standard. The reference range value is 31.3-4,000 pg/mL.
Use at an assay dependent concentration.
Uniprot #: SwissProt: P13597 Mouse
Alternative names: Antigen identified by monoclonal BB2 BB 2 BB2 CD 54 CD_antigen=CD54 CD54 Cell surface glycoprotein P3.58 Human rhinovirus receptor ICAM 1 ICAM-1 ICAM1 intercellular adhesion molecule 1 (CD54) intercellular adhesion molecule 1 (CD54), human rhinovirus receptor Intercellular adhesion molecule 1 Major group rhinovirus receptor MALA 2 MALA2 MyD 10 MyD10 P3.58 Surface antigen of activated B cells Surface antigen of activated B cells, BB2
Images
HA723833B_1.png Fig1: Sandwich ELISA analysis of Mouse ICAM1 matched pair antibodies

Capture: HA723832, Mouse ICAM1 Rabbit mAb [PSH16-34]
Detector: HA723833, Mouse ICAM1 Rabbit mAb [PSH16-35]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723832) 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 Mouse ICAM1 protein (HA211066) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA723833, 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.
HA723833B_2.png Fig2: Sandwich ELISA analysis of Mouse ICAM1 matched pair antibodies

Capture: HA723834, Mouse ICAM1 Rabbit mAb [PSH16-36]
Detector: HA723833, Mouse ICAM1 Rabbit mAb [PSH16-35]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723834) 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 Mouse ICAM1 protein (HA211066) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA723833, 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.
HA723833B_3.png Fig3: Interpolated concentrations of native ICAM-1 in mouse serum samples.

Capture: HA723832, Mouse ICAM1 Rabbit mAb [PSH16-34]
Detector: HA723833, Mouse ICAM1 Rabbit mAb [PSH16-35]

The concentrations of ICAM-1 were measured in duplicates, interpolated from the ICAM-1 standard curve and corrected for sample dilution. Undiluted samples are mouse serum 0.25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ICAM-1 concentration was determined to be 930.1 ng/ml in mouse serum.
HA723833B_4.png Fig4: Interpolated concentrations of native ICAM-1 in mouse serum samples.

Capture: HA723834, Mouse ICAM1 Rabbit mAb [PSH16-36]
Detector: HA723833, Mouse ICAM1 Rabbit mAb [PSH16-35]

The concentrations of ICAM-1 were measured in duplicates, interpolated from the ICAM-1 standard curve and corrected for sample dilution. Undiluted samples are mouse serum 0.125%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ICAM-1 concentration was determined to be 1,171 ng/ml in mouse serum.
HA723833B_5.png Fig5: Interpolated concentrations of native ICAM-1 in mouse LPS and mouse IFN-γ stimulated J774A.1 cell culture supernatant.

Capture: HA723832, Mouse ICAM1 Rabbit mAb [PSH16-34]
Detector: HA723833, Mouse ICAM1 Rabbit mAb [PSH16-35]

J774A.1 cells were stimulated with 1 µg/ml LPS and 100 ng/ml mouse IFN-γ or vehicle control and incubated for 72 hours. The concentrations of ICAM-1 measured in duplicate and interpolated from the ICAM-1 standard curve and corrected for sample dilution. Undiluted samples are as follows: stimulated 20%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ICAM-1 concentration was determined to be 6,529 pg/ml in LPS and mouse IFN-γ stimulated J774A.1 cell culture supernatant.
HA723833B_6.png Fig6: Interpolated concentrations of native ICAM-1 in mouse LPS and mouse IFN-γ stimulated J774A.1 cell culture supernatant.

Capture: HA723834, Mouse ICAM1 Rabbit mAb [PSH16-36]
Detector: HA723833, Mouse ICAM1 Rabbit mAb [PSH16-35]

J774A.1 cells were stimulated with 1 µg/ml LPS and 100 ng/ml mouse IFN-γ or vehicle control and incubated for 72 hours. The concentrations of ICAM-1 measured in duplicate and interpolated from the ICAM-1 standard curve and corrected for sample dilution. Undiluted samples are as follows: stimulated 20%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ICAM-1 concentration was determined to be 5,372 pg/ml in LPS and mouse IFN-γ stimulated J774A.1 cell culture supernatant.
HA723833B_7.png Fig7: Interpolated concentrations of spiked ICAM-1 in mouse cell culture media samples.

Capture: HA723832, Mouse ICAM1 Rabbit mAb [PSH16-34]
Detector: HA723833, Mouse ICAM1 Rabbit mAb [PSH16-35]

The concentrations of ICAM-1 were measured in duplicates, interpolated from the ICAM-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).
HA723833B_8.png Fig8: Interpolated concentrations of spiked ICAM-1 in mouse cell culture media samples.

Capture: HA723834, Mouse ICAM1 Rabbit mAb [PSH16-36]
Detector: HA723833, Mouse ICAM1 Rabbit mAb [PSH16-35]

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