Immunofluorescence analysis of Apolipoprotein A1 was performed 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Apolipoprotein A1 (311) Mouse Monoclonal Antibody (MIA1402) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified human plasma Apo A-1|
|Storage buffer||PBS, pH 7.4|
|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|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|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, MIA1402 can be used as a detection antibody with Product # 710263 or # 701239 as a coating antibody, to generate a matched pair. Using these matched pairs, recombinant human Apo A-1, but not recombinant mouse Apo A-1, was detected. MIA1402 can be used to detect Apo A-1 from serum samples. To increase sensitivity of sandwich ELISAs with MIA1402, a biotinylated detection antibody followed by Streptavidin-HRP is recommended.
By Western blot, MIA1402 detects recombinant human Apo A-1, but not recombinant mouse Apo A-1. MIA1402 may not be as successful for detecting endogenous Apo A-1 by Western blot.
MIA1402 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.