anti-NOS Clone 1E8-B8 antibody product blog
Tags: Antibody; Monoclonal Antibody; NOS Clone 1E8-B8; anti-NOS Clone 1E8-B8 antibody;
The NOS Clone 1E8-B8 n/a (Catalog #MBS350033) is an Antibody and is intended for research purposes only. The product is available for immediate purchase. The Anti-human inducible Nitric Oxide Synthase Monoclonal Antibody 1E8-B8 reacts with HumanCrossreactivity Note: Cross reacts with eNOS Maps to hiNOS (985-1002) and may cross-react with other species as described in the data sheet. MyBioSource\'s NOS Clone 1E8-B8 can be used in a range of immunoassay formats including, but not limited to, Western Immunoblot, Western Blot, Immunofluorescent Staining of Induced Cells.
Western Blot: Western immunoblots resulted in a single band being detected at ~ 130 kDa at a dilution of 1:50.
Immunofluorescent: This monoclonal antibody has been found to stain cells induced to produce iNOS at a 1:20 dilution. The ability of this monoclonal antibody to bind to iNOS in fixed cells was examined using three different cell lines. DLD-1(a human epithelial cell line). A-172 (a human glioblastoma cell line) and RAW 264.7 (a mouse macrophage cell line). The cells were cultured for 2 days in normal medium and then induced to produce iNOS by treatment for 16-24 hours with a cytokine/LPS mixture. Following the treatment. the cells were washed x 3 and fixed in either neutral buffered formalin or 100% ice cold acetone. They were reacted for 60 minutes with the culture supernatant. and then with FITC-conjugated goat anti-mouse IgG. The immunofluorescent staining pattern was observed using epifluorescent microscopy.
Immunofluorescent: This monoclonal antibody has been found to stain cells induced to produce iNOS at a 1:20 dilution. The ability of this monoclonal antibody to bind to iNOS in fixed cells was examined using three different cell lines. DLD-1(a human epithelial cell line). A-172 (a human glioblastoma cell line) and RAW 264.7 (a mouse macrophage cell line). The cells were cultured for 2 days in normal medium and then induced to produce iNOS by treatment for 16-24 hours with a cytokine/LPS mixture. Following the treatment. the cells were washed x 3 and fixed in either neutral buffered formalin or 100% ice cold acetone. They were reacted for 60 minutes with the culture supernatant. and then with FITC-conjugated goat anti-mouse IgG. The immunofluorescent staining pattern was observed using epifluorescent microscopy.
Western Blot Protocol: 1. After SDS-PAGE (on either 4-15% gradient gels or single percentage gels. such at 7.5% gels) and electrophoretic transfer to PVDF membrane. block the membrane overnight with 4% normal goat serum in TBS/Tween-20 buffer.
2. Wash x 2 with TBS/Tween-20.
3. Apply the mouse monoclonal antibody after preparing a 1:50 dilution. Use 2% normal goat serum in TBS/Tween-20 as buffer. and let the primary antibody bind for 2-4 hours.
4. Wash x 3 with TBS/Tween-20.
5. Apply affinity purified HRP-goat anti-rabbit IgG antiserum diluted 1:2500 (dilution may vary depending upon supplier) in 2% normal goat serum in TBS/Tween-20. Incubate 1-2 hours. Note: greater sensitivity may be achieved using ABC techniques.
6. Wash x 4 for 5 min per wash in TBS/Tween-20 buffer.
7. Develop color using the enhanced DAB reaction. Researchers should empirically determine the suitability of the NOS Clone 1E8-B8 n/a for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process.
The NOS Clone 1E8-B8 n/a product has the following accession number(s) (GI #295410244). Researchers may be interested in using Bioinformatics databases such as those available at The National Center for Biotechnology Information (NCBI) website for more information about accession numbers and the proteins they represent. Even researchers unfamiliar with bioinformatics databases will find the NCBI databases to be quite user friendly and useful.
To buy or view more detailed product information and pricing, please click on the technical datasheet page below:
This culture supernatant contains mouse monoclonal antibody clone 1E8-B8 raised against recombinant hiNOS. It has been epitope mapped using 96 overlapping 18 amino acid long synthetic peptides which cover the entire 1153 amino acid length of hiNOS and was found to bind to a defined region of the hiNOS sequence. residues 985-1002. This monoclonal antibody has been found to stain hiNOS and ecNOS in western immunoblots and by immunocytochemistry. This monoclonal antibody was tested for recognition of other NOS isoforms by ELISA. western immunoblotting. and immunocytochemical techniques. It has been found to be a mouse IgG1 kappa by isotyping.