Immunohistochemistry analysis of NF-kappa-B[p65] showing staining in the Cytoplasm and Nucleus of paraffin-embedded human breast carcinoma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a NF-kappa-B[p65] mouse monoclonal antibody (339900) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
|Tested species reactivity||Human|
|Published species reactivity||Human, Mouse, Not Applicable|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Recombinant protein (~175 amino acids) from the C-terminal portion of human NF-kappa-B (p65).|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||3 µg/ml|
|ELISA (ELISA)||Assay Dependent|
|Gel Shift (GS)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* 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.
NF-kappaB (Nuclear Factor kappa B) is a nuclear transcription factor found in all cell types and is involved in cellular responses to stimuli such as stress, cytokines, free radicals, ultraviolet irradiation, and bacterial or viral antigens. NF-kappaB plays a key role in regulating the immune response to infection. Consistent with this role, incorrect regulation of NF-kappaB has been linked to cancer, inflammatory and autoimmune diseases, septic shock, viral infection and improper immune development. There are five members in the NF-kappaB family: NF-kappaB1, NF-kappaB2, RelA (also named p65), RelB, and c-Rel. The most common form of NF-kappaB is the p50/RelA heterodimer, although other forms of NF-kappaB dimers, such as p50/p50, p52/p52, p52/RelA, p50/c-Rel, c-Rel/c-Rel, p52/RelB, and p50/RelB, have also been identified in some types of cells. The primary role of NF-kappaB is to maintain normal cellular functions that range from cell-to-cell communication to cell motility, cell cycle progression, and cell lineage development. The activity of NF-kappaB is tightly regulated by interaction with inhibitory IkappaB proteins.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Human||Not Cited||Antipsychotic drugs suppress the AKT/NF-¿B pathway and regulate the differentiation of T-cell subsets.||Chen ML,Tsai TC,Lin YY,Tsai YM,Wang LK,Lee MC,Tsai FM||Immunology letters (140:81)||2011|
Cautionary notes on the use of NF-¿B p65 and p50 antibodies for CNS studies.
33-9900 was used in immunocytochemistry, immunohistochemistry (free floating), and western blot to examine specificity of the anti-NF-κB antibodies.
|Herkenham M,Rathore P,Brown P,Listwak SJ||Journal of neuroinflammation (8:null)||2011|
|Not Applicable||Not Cited||
Sequence characteristics of functional siRNAs.
33-9900 was used in immunocytochemistry to develop an algorithm to generate effective siRNAs
|Jagla B,Aulner N,Kelly PD,Song D,Volchuk A,Zatorski A,Shum D,Mayer T,De Angelis DA,Ouerfelli O,Rutishauser U,Rothman JE||RNA (New York, N.Y.) (11:864)||2005|