Immunohistochemistry was performed on cancer biopsies of deparaffinized Human colon carcinoma tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing alpha Actinin 4 (MA1-800) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG|
|Immunogen||Synthetic peptide corresponding to residues A(884) P Y Q G P D A V P G A L D(897) of human alpha Actinin 4.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Flow Cytometry (Flow)||1µg per 10^6 cells|
|Immunofluorescence (IF)||4 ug/mL|
|Immunohistochemistry (Paraffin) (IHC (P))||4 µg/ml|
|Western Blot (WB)||2 ug/mL|
* 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.
|Western Blot (WB)||See 1 publications below|
MA1-800 detects the alpha Actinin 4 in human and mouse cells.
MA1-800 has been successfully used in Western blot, and immunofluorescence procedures. MA1-800 detects alpha Actinin 4 which has a predicted molecular weight of approximately 105 kDa. Under reducing conditions, MA1-800 detects a monomer of alpha Actinin 4 at approximately 52.5 kDa.
The MA1-800 immunogen is a synthetic peptide corresponding to residues A(884) P Y Q G P D A V P G A L D(897) of human alpha Actin 4.
alpha-Actinin 4, a member of the alpha-Actinin family of cytoskeletal proteins, functions as an actin-filament crosslinking protein. It has been shown to play a role in cell motility, endocytosis, and glomerular function.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Cell-surface processing of the metalloprotease pro-ADAMTS9 is influenced by the chaperone GRP94/gp96.
MA1-800 was used in western blot to show that pro-ADAMTS9 associates with heat shock protein gp96 at the cell surface
|Koo BH,Apte SS||The Journal of biological chemistry (285:197)||2010|