Human IgE Antibody (Fc-2001)

Western Blot: Human IgE Antibody (Fc-2001) [NBP1-78601] - Analysis of 0.5ug of purified human IgE, IgM, IgA and IgG per well and 10ul of PageRuler Prestained Protein Ladder per well.

Direct ELISA analysis of human IgE was performed by coating wells of a 96-well plate with 100ul per well of human IgE, IgM, IgA, or IgG, diluted incarbonate/bicarbonate buffer at concentrations rangingfrom 5-5000ng/ml, and incubating overnight at 4C. The plate was washedand blocked with StartingBlock T20 (TBS) Blocking Buffer (Product # 37543)for at least 30 minutes at room temperature, and then incubated with 100ulper well of a human IgE monoclonal antibody at aconcentration of 1ug/ml for 1 hour at room temperature. The plate waswashed, and incubated with 100ul per well of an HRP-conjugated goat antimouse IgG Fc secondary antibody at a dilution of 1:8000for at least 30 minutes at room temperature. Detection was performed byadding 100ul per well of 1-Step Ultra TMB substrate andincubating for a few minutes at room temperature. The reactions were thenstopped with TMB Stop Solution. Absorbances were readon a spectrophotometer at 450-550nm.

Sandwich ELISA analysis of human IgE was performed by coating wells ofa 96-well plate with100ul per well of a goat anti-human IgG (H+L) antibody diluted to a concentration of 1ug/ml incarbonate/bicarbonate buffer, and incubating overnight at4C. The plate was washed and blocked with StartingBlock T20 (TBS)Blocking Buffer for at least 30 minutes at roomtemperature, and then 100ul per well of human IgE, IgM, IgA, or IgG, dilutedin carbonate/bicarbonate buffer at concentrations rangingfrom 1-1000ng/ml, was added to the plate and incubated for at least 1 hourat room temperature. The plate was washed with 150ul per well of 1X TBST,and 100ul per well of a human IgE monoclonal antibody was added to the wells at a concentration of 1ug/ml, and incubatedfor 1 hour at room temperature. The plate was washed, and incubated with100ul per well of an HRP-conjugated goat anti-mouse IgG Fc secondaryantibody at a dilution of 1:8000 for at least 30 minutes atroom temperature. Detection was performed by adding 100ul per well of 1-Step Ultra TMB substrate and incubating for a few minutesat room temperature. The reactions were then stopped with TMB StopSolution . Absorbances were read on a spectrophotometerat 450-550nm.

Sandwich ELISA analysis of human IgE was performed by coating wells ofa 96-well plate with100ul per well of a goat anti-human IgG (H+L) antibody diluted to a concentration of 1ug/ml incarbonate/bicarbonate buffer, and incubating overnight at4C. The plate was washed and blocked with Starting block T20 (TBS)Blocking Buffer for at least 30 minutes at roomtemperature, and then 100ul per well of human IgE, diluted in 1X PBS atconcentrations ranging from 1-1000ng/ml (left panel), and normal humanserum or serum from an asthma patient on anti-asthma prescriptionmedications (control serum, right panel), diluted in 1X PBS at dilutionsranging from 1:5 to 1:5000, was added to the plate and incubated for at least1 hour at room temperature. The plate was washed with 150ul per well of1X TBST, and 100ul of a human IgE monoclonal antibody was added to the wells at a concentration of 1ug/ml, and incubatedfor 1 hour at room temperature. The plate was washed and incubated with100ul per well of an HRP-conjugated goat anti-mouse IgG Fc secondaryantibody at a dilution of 1:8000 for at least 30 minutes atroom temperature. Detection was performed by adding 100ul per well of 1-Step Ultra TMB substrate and incubating for a few minutesat room temperature. The reactions were then stopped with TMB StopSolution. Absorbances were read on a spectrophotometerat 450-550nm.

• Reactivity: Hu
• Applications: WB, ELISA, Func, IP
• Clone: Fc-2001
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

Human IgE Antibody (Fc-2001) Summary

• Immunogen: Human Myeloma IgE
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Purity: Ion exchange chromatography

Applications/Dilutions

• Dilutions:
  • ELISA 1:100 - 1:2000
  • Immunoprecipitation 1:10 - 1:500
  • Radioimmunoassay 1:100 - 1:2000
  • Western Blot 1:500 - 1:2000

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS, potassium phosphate with 1mg/ml BSA, and 50% glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Ion exchange chromatography

Alternate Names for Human IgE Antibody (Fc-2001)

• immunoglobulin heavy constant epsilon

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.