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mAbs Against MOUSE Protein

SAA3

Antibody Validated
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
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
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) (+)