Additional information
Specifity
Using Nitroso-L-Cysteine-BSA conjugate (NO-Cys-BSA), antibody specificity was performed by using an ELISA test by competition experiments:
| Nitroso-L-Cysteine-BSA | Kd around 10-8 M | very high affinity and specificity |
Low affinity or no crossreactivity with close competitors:
| Competitors | Affinity compared to NO-Cys-BSA | |
|---|---|---|
| L-Cysteine-BSA | 200 times lower | low affinity |
| Nitroso-N-Acetyl-Cysteine-BSA | 50 000 times lower | no crossreactivity |
| N-Acetyl-Cysteine-BSA | 50 000 times lower | no crossreactivity |
The antibody recognizes the conjugated form of the small molecule, not the free form (see protocol for specific tissue preparation).
Additional Information
- Description
Monoclonal antibody was obtained after BALB/c mouse immunisation with the conjugate: Nitroso-Cysteine-bovine serum albumin (BSA). Hybridoma clone was obtained by hybridization of spleen cells with the myeloma cell line SP2/O/Ag14
- Host species
Mouse
- Clonality
Monoclonal
- Clone number
HY8E12
- Sub-categories
Nitrosylated compounds
- Isotype
IgG, Kappa
- Target
Conjugated Nitroso-L-Cysteine
- Immunogen
Nitrosylated Synthetic L-Cysteine conjugated to bovine serum albumin (BSA)
- Species reactivity
The antibody recognizes the target in all species, vertebrates (included humans) and invertebrates, because of the universality of the small molecule
- Purity
Monoclonal antibody was preadsorbed on protein carriers and purified by ammonium sulfate precipitation
- Form
Lyophilized
- Stabilizers / preservatives
None
- Storage instructions and buffer
Lyophilized vial must be stored at 4°C in a dry area up to 2 years after receipt. After reconstitution with 50% of distilled water and 50% of glycerol, the aliquot must be stored at -20°C, and is stable at least for 2 years. Avoid freeze-thaw cycles
- Research areas
Biochemistry, Immunology, Neurobiology, Neurodegenerative diseases, Pharmacology
- Applications
ELISA, Immunohistochemistry (Glutaraldehyde and PFA fixation, see protocol), Immunocytochemistry. Optimal dilutions should be determined by each laboratory for each application. Suggested starting dilutions: 1/2000 – 1/10000 for ELISA, 1/1000 – 1/5000 for IHC
- Corresponding antigen/related product
To validate the specificity of the response in your biological samples, Polyneuros sells as well the corresponding antigen: Nitroso-L-Cysteine conjugate (Product number: AG122)
- Related antibodies
Polyneuros also sells polyclonal antibody against another small molecule involved in the oxidative stress: anti-Malondialdehyde (AP050)