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Western blot analysis was performed on whole cell extracts (30 µg lysate) of Daudi (Lane 1), K-562 (Lane 2), NIH/3T3 (Lane3), SH-SY5Y (Lane4) and tissue extract (30 µg lysate) of Rat Testis (Lane 5).The blots were probed with Anti-DNA Polymerase beta Mouse monoclonal Antibody (Product # MA5-12086, 2 µg/ml) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjµgate (Product # A28177, 0.4 µg/ml, 1:2500 dilution). A 38 kDa band corresponding to DNA Polymerase beta was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). 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||Human, Mouse, Rat|
|Published species reactivity||Human, Mouse|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||DNA polymerase beta protein|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Western Blot (WB)||1-3 µg/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 2 publications below|
MA5-12086 targets DNA Polymerase beta in WB applications and shows reactivity with Human samples.
The MA5-12086 immunogen is dNA polymerase beta protein.
DNA polymerase beta comprises an amino-terminal 8-kDa domain and a carboxy-terminal 31-kDa domain. The N-terminal ssDNA binding domain has a deoxyribose phosphodiesterase activity while the C-terminal domain has a nucleotidyltransferase activity. Mammalian DNA polymerase beta, a DNA repair polymerase, is constitutively expressed in cultured cells, but treatment of cells with the DNA-alkylating agents such as N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or methyl methanesulfonate up-regulates beta-pol level. DNA polymerase beta fills single nucleotide gaps in DNA produced by the base excision repair pathway of mammalian cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Transcriptional profiling reveals elevated Sox2 in DNA polymerase ß null mouse embryonic fibroblasts.
MA5-12086 was used in western blot to study the upregulation of Sox2 gene expression in murine embryonic fibroblasts deleted for DNA polymerase beta
|Li J,Luthra S,Wang XH,Chandran UR,Sobol RW||American journal of cancer research (2:699)||2012|
Parp1 activation in mouse embryonic fibroblasts promotes Pol beta-dependent cellular hypersensitivity to alkylation damage.
MA5-12086 was used in western blot to investigate the effect of Parp1 on cellular hypersensitivity to alkylation damage
|Jelezcova E,Trivedi RN,Wang XH,Tang JB,Brown AR,Goellner EM,Schamus S,Fornsaglio JL,Sobol RW||Mutation research (686:57)||2010|
DNA pol beta; DNA polymerase beta; DNA polymerase beta subunit; DNA-directed DNA polymerase beta; Pol beta; polymerase (DNA directed), beta; polymerase (DNA) beta; polymerase-beta