Immunohistochemistry analysis of Apolipoprotein A-1 showing staining in the cytoplasm of paraffin-embedded human liver tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Apolipoprotein A-1 Mouse Monoclonal Antibody (MIA1401) diluted in 3% BSA-PBS at a dilution of 1:100 for 1 hour at 37ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Purified human plasma Apo A-1|
|Storage buffer||PBS, pH 7.2|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100|
|Immunoprecipitation (IP)||Assay Dependent|
|Radioimmune Assays (RIA)||Assay Dependent|
|Western Blot (WB)||1:500-1:2000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
By sandwich ELISA, MIA1401 can be used as a detection antibody with Product # 710263 as the coating antibody, to generate a matched pair. Using this matched pair, recombinant human Apo A-1, but not recombinant mouse Apo A-1, was detected. It is not recommended to use Product # 701239 for coating when MIA1401 is used for detection.MIA1401 can be used to detect Apo A-1 from serum samples. To increase sensitivity of sandwich ELISAs with MIA1401, a biotinylated detection antibody followed by Streptavidin-HRP is recommended.
By Western blot, MIA1401 detects recombinant human Apo A-1, but not recombinant mouse Apo A-1. MIA1401 may not be as successful for detecting endogenous Apo A-1 by Western blot.
MIA1401 was formerly sold as a Seradyn product.
This gene encodes apolipoprotein A-I, which is the major protein component of high density lipoprotein (HDL) in plasma. The protein promotes cholesterol efflux from tissues to the liver for excretion, and it is a cofactor for lecithin cholesterolacyltransferase (LCAT) which is responsible for the formation of most plasma cholesteryl esters. This gene is closely linked with two other apolipoprotein genes on chromosome 11. Defects in this gene are associated with HDL deficiencies, including Tangier disease, and with systemic non-neuropathic amyloidosis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.