Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), Hep G2 (Lane 2), SK-OV-3 (Lane 3), COS-7 (Lane 4), NIH/3T3 (Lane 5), C2C12 (Lane 6), PC-3 (Lane 7), and LNCaP (Lane 8). The blots were probed with Rabbit Anti-PEBP1 Polyclonal Antibody (Product # PA3211, 1:250 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 ug/ml, 1:4000 dilution). A 21 kDa band corresponding to PEBP1 was observed across the cell lines tested. The 42 kDa bands observed could potentially be PEBP1 dimers, caused by the PKC-mediated phosphorylation of PEBP1, as reported in literature. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), 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, Non-human primate, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to the C-terminal amino acids C(167)YQAEWDDYVPKLYEQLSGK(187) of PEBP1|
|Storage buffer||whole serum|
|Contains||0.02% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:2000|
|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.
PA3-211 detects PEBP1 in human, mouse, and rat samples.
PA3-211 has successfully been used in Western blot, immunocytochemical, and immunohistochemical (paraffin) procedures.
The PA3-211 immunogen is a synthetic peptide corresponding to the C-terminal amino acids C(167)YQAEWDDYVPKLYEQLSGK(187) of PEBP1
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.