mAbs Against MOUSE Protein
SAA3
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas |
| STATUS: | Validated |
| TYPE: | Rat anti mouse |
| CLONE NAME: | JOR110A |
| PROTEIN: | Serum amyloid A-3 protein precursor |
| PROTEIN WEB: | http://www.ncbi.nlm.nih.gov/protein/6755396 |
| ANTIGEN USED: | His-mSAA3 (20-122aa) recombinant protein |
| FUSION PARTNER: | NS1/Ag4-1 (NS1) cells |
| ISOTYPE: | iGg2b |
| SPECIES REACTIVITY: | mouse |
| PREPARATION AND STORAGE: | Aliquot and store at 4C. Do not freeze |
Description
Serum amyloid A (SAA) proteins are proposed mediators of inflammation and metabolism, with increased serum levels being associated with obesity, chronic hyperglycemia, insulin resistance and cardiovascular disease. Mouse SAA3 (mSAA3) proten is known to be up-regulated extrahepatically in inflammatory responses, and acts as an endogenous ligand for the toll-like receptor 4/MD-2 complex. SAA3 also displays monocyte chemotactic activity and may play a role in metabolic inflammation.
Applications
| IHC Techniques | Clone | Dilution | Antibody concentration | Antigen retrieval method | Visualization kit | +/- control | Protein localization | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|---|
| Frozen tissue and cytospins | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | JOR110A | Neat supernatant | / | ||||||
| Paraffin tissue | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | JOR110A | Neat supernatant | Ventana | / | |||||
| Immunofluorescence | |||||||||
Enlarge image
- JOR110A in HEK293T transfected cells.
- To confirm that JOR110A mAb recognized mSAA3 protein, immunohistochemistry on frozen cytospins preparations of MYC-tagged mSAA3 expressed in HEK 293T cell line was performed. Cytospin preparation of HEK 293T-MYC-mSAA1, HEK 293T-MYC-mSAA2 and HEK 293T-MYC-mSAA4 transfected cells were used to confirm the specificity of the antibody.
Enlarge image
- JOR110A mAb in mouse paraffin tissues.
- JOR110A antibody can be used to detect mSAA3 protein in mouse paraffin embedded tissue samples.
| WB Techniques | Clone | Dilution | Antibody concentration | +/- control | Expected MW | Observed Mw | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|
| Western Blotting | ||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | JOR110A | Neat supernatant | / | 13kDa | 13kDa | Available | ||
| Immunoprecipitation | ||||||||
Enlarge image
- JOR110A mAb in Western blotting technique
- Since mSaa3 is a secreted protein we have collected and concentrated the supernatant from the RAW12 cell line (mouse macrophage) unstimulted (Neg control) and stimulated with LPS after 8, 12, 16 and 24 hours as reported in the attached article. A specific band of 13kDa has been deteced in all actived LPS RAW12 cell lines, with the higher expression after 12 and 16 hour stimulation. No mSaa3 protein has been decected in the concentrated supernatant of the RAW12 cell line unstimulated.
Lane 1 RAW supernatant after 8 hours (20ul) (+)
Lane 2 RAW supernatant after 12 hours (20ul) (+)
Lane 3 RAW supernatant after 16 hours (20ul) (+)
Lane 4 RAW supernatant after 24 hours (20ul) (+)
Lane 5 RAW supernatant unstimulated (20ul) (-)
Lane 6 HEK 293T-GFP-mSAA3 trasfected cells (20ug) (+)
Enlarge image
- JOR110A mAb in Western blotting technique
- Lane 1 HEK-mSAA1 transfected cells (10ug) (-)
Lane 2 HEK-mSAA2 transfected cells (10ug) (-)
Lane 3 HEK-mSAA3 transfected cells (10ug) (+)
Lane 4 HEK-mSAA4 transfected cells (10ug) (-)
Lane 5 RAW supernatant unstimulated (20ul) (-)
Lane 6 RAW supernatant stimulated 12h (20ul) (+)