Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- ELISA [2]
Submit
Validation data
Reference
Comment
Report error
- Product number
- LS-C355485 - Provider product page
- Provider
- LSBio
- Product name
- IL6 / Interleukin 6 Antibody LS-C355485
- Antibody type
- Polyclonal
- Description
- Protein G purified
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Storage
- Store at -20°C. Avoid freeze-thaw cycles.
No comments: Submit comment
Supportive validation
- Submitted by
- LSBio (provider)
- Enhanced method
- Genetic validation
- Main image
- Experimental details
- Western blot analysis of IL-6 was performed by loading 20µg of LPS-treated THP-1 whole cell lysates in non-reducing sample buffer and 8ul PageRuler Plus Prestained Protein Ladder per well onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter and blocked with 5% Milk/TBST for at least 1 hour at room temperature. IL-6 was detected using a IL-6 rabbit polyclonal antibody at a dilution of 1:1000 in blocking buffer overnight at 4°C on a rocking platform, followed by a HRP conjugated secondary antibody at a dilution of 1:5000 for at least 1 hour at room temperature. Chemiluminescent detection was performed using SuperSignal West Dura Extended Duration Substrate and the myECL Imager.
Supportive validation
- Submitted by
- LSBio (provider)
- Enhanced method
- Genetic validation
- Main image
- Experimental details
- Direct ELISA analysis of LPS-treated IL-6 was performed by coating wells of a 96-well plate with 100µl per well of IL-6 diluted in carbonate/bicarbonate buffer at a concentration of 2 µg/ml overnight at 4C. Wells of the plate were washed, blocked with starting blocking buffer, and incubated with 100ul per well of a rabbit anti-IL-6 oligoclonal antibody at a serially dilution of 1:50, 1:100, 1:250, 1:500, 1:1000, 1:2000 and 1:5000 for 90 minutes at 37C. The plate was washed, then incubated with 100µl per well of an HRP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:5000 for 90 minutes at 37C. Detection was performed using 1-Step Ultra TMB substrate for 5-10 minutes at room temperature in the dark. The reaction was stopped with Stop solution, and absorbances were read on a spectrophotometer at 450-550nm.
- Submitted by
- LSBio (provider)
- Main image
- Experimental details
- Direct ELISA analysis of LPS-treated IL-6 was performed by coating wells of a 96-well plate with 100µl per well of IL-6 diluted in carbonate/bicarbonate buffer at a concentration of 2 µg/ml overnight at 4C. Wells of the plate were washed, blocked with starting blocking buffer, and incubated with 100ul per well of a rabbit anti-IL-6 oligoclonal antibody at a serially dilution of 1:50, 1:100, 1:250, 1:500, 1:1000, 1:2000 and 1:5000 for 90 minutes at 37C. The plate was washed, then incubated with 100µl per well of an HRP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:5000 for 90 minutes at 37C. Detection was performed using 1-Step Ultra TMB substrate for 5-10 minutes at room temperature in the dark. The reaction was stopped with Stop solution, and absorbances were read on a spectrophotometer at 450-550nm.