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Immunohistochemistry was performed on human liver tissue sections. To expose Apo A-1, tissues were microwaved for 10 minutes in 10mM sodium citrate, pH 6.0. Following antigen retrieval, endogenous peroxidases were blocked in a 3% hydrogen peroxide-methanol solution for 15 minutes, and then tissues were blocked in 3% BSA in PBS for 30 minutes at room temperature. Tissues were probed with an Apo A-1 monoclonal antibody (Product # MA5-14667) at a dilution of 1:100 overnight at 4C, washed extensively in PBST, and detection was performed with an HRP-conjugated anti-mouse IgG secondary antibody followed by DAB substrate. Tissues were counterstained with hematoxylin.
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Purified human plasma Apo A-1|
|Storage buffer||0.01M potassium phosphate with 0.135M NaCl, 2.5mM KCl, 1mg/ml BSA|
|Contains||0.05% 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 *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50-1:200|
|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, MA5-14667 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 MA5-14667 is used for detection. By sandwich ELISA, MA5-14667 can be used to detect Apo A-1 from serum samples. To increase sensitivity of sandwich ELISAs with MA5-14667, a biotinylated detection antibody followed by Streptavidin-HRP is recommended.
By Western blot, MA5-14667 detects recombinant human Apo A-1, but not recombinant mouse Apo A-1. MA5-14667 may not be as successful for detecting endogenous Apo A-1 by Western blot.
Product MA514667 is a smaller package size of MIA1401 (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.