Western blot analysis was performed on nuclear extracts (30µg lysate) of A-431 (Lane 1), MCF7 (Lane 2), LNCap (Lane 3), DU 145 (Lane 4), HeLa (Lane 5) and U-2 OS (Lane 6). The blots were probed with anti-MBTD1 Mouse Monoclonal Antibody (Product# 730065, 2µg/ml) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25µg/ml, 1:4000 dilution). A 70 kDa band corresponding to MBTD1 was observed across 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 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. This is followed by incubating the membrane with Poly-HRP Streptavidin (Product# N200, 1:10,000 dilution). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||130-566 of 628 of MBTD1 Protein.|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||2 ug|
|Western Blot (WB)||2 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.
|Immunoprecipitation (IP)||See 1 publications below|
MBDT1 is a putative Polycomb group (PcG) protein. PcG proteins maintain the transcriptionally repressive state of genes, probably via a modification of chromatin, rendering it heritably changed in its expressibility. MBDT1 specifically binds to monomethylated and dimethylated 'Lys-20' on histone H4.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Assessment of a method to characterize antibody selectivity and specificity for use in immunoprecipitation.
730065 was used in immunoprecipitation to describe a mass spectrometry-based method for scoring immunoprecipitation antibody quality
|Marcon E,Jain H,Bhattacharya A,Guo H,Phanse S,Pu S,Byram G,Collins BC,Dowdell E,Fenner M,Guo X,Hutchinson A,Kennedy JJ,Krastins B,Larsen B,Lin ZY,Lopez MF,Loppnau P,Miersch S,Nguyen T,Olsen JB,Paduch M,Ravichandran M,Seitova A,Vadali G,Vogelsang MS,Whiteaker JR,Zhong G,Zhong N,Zhao L,Aebersold R,Arrowsmith CH,Emili A,Frappier L,Gingras AC,Gstaiger M,Paulovich AG,Koide S,Kossiakoff AA,Sidhu SS,Wodak SJ,Gräslund S,Greenblatt JF,Edwards AM||Nature methods (12:725)||2015|