Immunofluorescence analysis of Apolipoprotein A1 was performed using 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 (513) Mouse Monoclonal Antibody (MIA1404) 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|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified human plasma Apo A-1|
|Storage buffer||0.015M potassium phosphate, pH 7.2, with 0.15M NaCl|
|Contains||0.1% 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.
|Immunohistochemistry (IHC)||See 1 publications below|
By sandwich ELISA, MIA1404 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. MIA1404 can be used to detect Apo A-1 from serum samples. To increase sensitivity of sandwich ELISAs with MIA1404, a biotinylated detection antibody followed by Streptavidin-HRP is recommended.
By Western blot, MIA1404 detects recombinant human Apo A-1, but not recombinant mouse Apo A-1. MIA1404 is also recommended for detecting endogenous Apo A-1 by Western blot.
MIA1404 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.
Apolipoprotein A-I and B distribution in the human cornea.
MIA1404 was used in immunohistochemistry to study the expression of apoliprotein A1 and B in human cornea
|Ashraf F,Cogan DG,Kruth HS||Investigative ophthalmology and visual science (34:3574)||1993|