Note: You clicked on an external link, which has been disabled in order to keep your shopping session open.
Western blot analysis was performed on whole cell extracts (30 ug lysate) of Jurkat (Lane 1), Jurkat treated with TNF alpha (Lane 2) . The blots were probed with ABfinity™ Rabbit Monoclonal Anti- NFkB p50 [pS337] Antibody (Product# 710460, 1-2 ug/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 ug/ml, 1:2500 dilution). A 55 kDa band corresponding to NFkB p50 [pS337]was observed across cell lines tested. 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|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Peptide corresponding to amino acids 333–341 of human p50 [pS337]|
|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 *|
|ChIP assay (ChIP)||1-3 ug x 10^6 cells|
|Western Blot (WB)||1-2 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.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale and purified with Protein A.
ABfinity™ oligoclonal antibodies comprise a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds—the sensitivity of a polyclonal antibody with the specificity of a monoclonal, all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody—recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets when compared with monoclonal antibodies—an oligoclonal antibody has a known mixture of light and heavy chains. This exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
The p50 protein is a member of the NF- kappaB/Rel transcription factor family that is a subunit of the NF- kappaB protein complex. p50 is processed from a 105 kDa precursor protein by the 26S proteasome, and can exist as either a heterodimer or a homodimer. Heterodimers formed with Rel proteins activate transcription, but because p50 lacks a C terminal transactivation domain, the homodimer acts as a repressor unless it associates with a co-activator such as BCL3. The NF- kappaB protein complex expressed in most cell types, and is primarily found in the cytoplasm in an inactive state in association with I-kappa-B. Phosphorylation of I-kappa-B releases the complex, allowing translocation to the nucleus and modification of gene expression. Phosphorylation of the p50 subunit at the Ser-337 residue has been shown to significantly increase DNA binding efficiency of the NF- kappaB protein complex.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
DNA binding factor KBF1; DNA-binding factor KBF1; NF-kappabeta; nuclear factor kappa-B DNA binding subunit; nuclear factor NF-kappa-B p105 subunit; nuclear factor NF-kappa-B p50 subunit; nuclear factor of kappa light polypeptide gene enhancer in B-cells 1; OTTHUMP00000219572
CVID12; EBP-1; KBF1; NF-kappa-B; NF-kappaB; NF-kB1; NFkappaB; NFKB-p105; NFKB-p50; NFKB1; p105; p50