Western blot analysis was performed on whole cell extracts (30 ug lysate) of HeLa (Lane 1), Jurkat (Lane 2), and tissue extracts (30 µg lysate) of Mouse testis (Lane 3), Mouse pancreas (Lane 4), and Rat pancreas (Lane 5). The blot was probed with Anti-PSME4 Rabbit Polyclonal Antibody (Product # PA1-1961, 1:250 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 200 kDa band corresponding to PSME4 was observed in HeLa, Jurkat, and mouse testis. In addition, a 140 kDa band corresponding to an isoform of PSME4 was observed across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and HiMark™ Pre-stained Protein Standard (Product # LC5699). Resolved proteins were then transferred onto a nitrocellulose membrane using the overnight wet transfer (tank transfer) technique. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Bovine, Human, Mouse, Rat|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues T(1620) K L P K K R K R D P G S V G(1634) of human PA200.|
|Storage buffer||whole serum|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:250|
* 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.
|Miscellaneous PubMed (MISC)||See 1 publications below|
PA1-1961 detects PA200 from bovine, human, mouse and rat samples.
PA1-1961 has been successfully used in Western blot, immunofluorescence and immunoprecipitation procedures. By Western blot, this antibody detects an ~200 kDa protein representing PA200 from bovine testis tissue extract. In mouse tissues, this antibody detects both a 200 kDa and a 160 kDa protein. In mouse tissues, the 200 kDa species is most abundant in testis whereas the 160 kDa species is enriched in the spleen. Immunofluorescence staining of PA200 in HeLa cells results in diffuse nuclear staining prior to ionizing radiation treatment. Immunofluorescence staining of PA200 in HeLa cells after ionizing radiation treatment results in focal aggregate staining within the nucleus.
The PA1-1961 immunogen is a synthetic peptide corresponding to residues T(1620) K L P K K R K R D P G S V G(1634) of human PA200.
The 26S proteasome is a multicatalytic proteinase complex with a highly ordered structure composed of 2 complexes, a 20S core and a 19S regulator. The 20S core is composed of 4 rings of 28 non-identical subunits; 2 rings are composed of 7 alpha subunits and 2 rings are composed of 7 beta subunits. The 19S regulator is composed of a base, which contains 6 ATPase subunits and 2 non-ATPase subunits, and a lid, which contains up to 10 non-ATPase subunits. Proteasomes are distributed throughout eukaryotic cells at a high concentration and cleave peptides in an ATP/ubiquitin-dependent process in a non-lysosomal pathway. An essential function of a modified proteasome, the immunoproteasome, is the processing of class I MHC peptides. The immunoproteasome contains an alternate regulator, referred to as the 11S regulator or PA28, that replaces the 19S regulator. Three subunits (alpha, beta and gamma) of the 11S regulator have been identified. This gene encodes the alpha subunit of the 11S regulator, one of the two 11S subunits that is induced by gamma-interferon. Three alpha and three beta subunits combine to form a heterohexameric ring. Two transcripts encoding different isoforms have been identified.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
PA200, a nuclear proteasome activator involved in DNA repair.
PA1-1961 was used in immunohistochemistry and western blot to identify a nuclear proteasome activator DNA repair protein PA200
|Ustrell V,Hoffman L,Pratt G,Rechsteiner M||The EMBO journal (21:3516)||2002|