Tested Applications
| Positive WB detected in | HeLa cells, HEK-293 cells, MOLT-4 cells, Jurkat cells, Raji cells |
| Positive IHC detected in | human breast cancer tissue Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0 |
Recommended dilution
| Application | Dilution |
|---|---|
| Western Blot (WB) | WB : 1:1000-1:4000 |
| Immunohistochemistry (IHC) | IHC : 1:150-1:600 |
| It is recommended that this reagent should be titrated in each testing system to obtain optimal results. | |
| Sample-dependent, Check data in validation data gallery. | |
Product Information
66535-1-Ig targets NF-κB p65 in WB, IHC, IF, CoIP, ChIP, RIP, ELISA applications and shows reactivity with human samples.
| Tested Reactivity | human |
| Cited Reactivity | human, pig, chicken, bovine, hamster |
| Host / Isotype | Mouse / IgG1 |
| Class | Monoclonal |
| Type | Antibody |
| Immunogen |
CatNo: Ag1199 Product name: Recombinant human p65; RELA protein Source: e coli.-derived, PGEX-4T Tag: GST Domain: 1-220 aa of BC011603 Sequence: MDELFPLIFPAEPAQASGPYVEIIEQPKQRGMRFRYKCEGRSAGSIPGERSTDTTKTHPTIKINGYTGPGTVRISLVTKDPPHRPHPHELVGKDCRDGFYEAELCPDRCIHSFQNLGIQCVKKRDLEQAISQRIQTNNNPFQVPIEEQRGDYDLNAVRLCFQVTVRDPSGRPLRLPPVLSHPIFDNRAPNTAELKICRVNRNSGSCLGGDEIFLLCDKVQ Predict reactive species |
| Full Name | v-rel reticuloendotheliosis viral oncogene homolog A (avian) |
| Calculated Molecular Weight | 65 kDa |
| Observed Molecular Weight | 65 kDa |
| GenBank Accession Number | BC011603 |
| Gene Symbol | NF-κB p65 |
| Gene ID (NCBI) | 5970 |
| RRID | AB_2881898 |
| Conjugate | Unconjugated |
| Form | Liquid |
| Purification Method | Protein G purification |
| UNIPROT ID | Q04206 |
| Storage Buffer | PBS with 0.02% sodium azide and 50% glycerol, pH 7.3. |
| Storage Conditions | Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage. 20ul sizes contain 0.1% BSA. |
Background Information
Nuclear factor k B (NF-kB) is a sequence-specific DNA-binding protein complex which regulates the expression of viral genomes, including the human immunodeficiency virus, and a variety of cellular genes, particularly those involved in immune and inflammatory responses. The members of the NF-Kb family in mammalian cells include the proto-oncogene c-Rel,p50/p105 (NFkB1), p65 (RelA), p52/p100 (NFkB2), and RelB. All of these proteins share a conserved 300-amino acid region known as the Rel homology domain which is responsible for DNA binding, dimerization, and nuclear translocation of NF-Kb. The p65 subunit is a major component of NF-Kb complexes and is responsible for trans-activation. NF-kappa-B heterodimeric p65-p50 and p65-c-Rel complexes are transcriptional activators. The NF-kappa-B p65-p65 complex appears to be involved in invasin-mediated activation of IL-8 expression. The inhibitory effect of I-kappa-B upon NF-kappa-B the cytoplasm is exerted primarily through the interaction with p65. p65 shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex. It associates with chromatin at the NF-kappa-B promoter region via association with DDX1.
Protocols
| Product Specific Protocols | |
|---|---|
| IF protocol for NF-κB p65 antibody 66535-1-Ig | Download protocol |
| IHC protocol for NF-κB p65 antibody 66535-1-Ig | Download protocol |
| WB protocol for NF-κB p65 antibody 66535-1-Ig | Download protocol |
| Standard Protocols | |
|---|---|
| Click here to view our Standard Protocols |
Publications
What published studies show
Anti-NF-κB p65 antibody (Cat# 66535-1-Ig) has accumulated 390 citations. Publications appear in journals including Nature Communications, Science Translational Medicine, Journal of Clinical Investigation, Advanced Science, Cell Reports, Oncogene, Theranostics, Biomaterials, and numerous others spanning immunology, oncology, neuroscience, pharmacology, and materials science. Research fields encompass inflammation, cancer, neurodegeneration, cardiovascular disease, hepatic/renal injury, traditional Chinese medicine, nanomedicine, oxidative stress, and metabolic disorders.
| Species | Application | Title |
|---|---|---|
Cell Mol Immunol SPT6 maintains epidermal homeostasis by inhibiting an NF-κB-positive feedback loop to prevent excessive inflammation. | ||
Cell Host Microbe Oat-β-glucan potentiates anti-PD-1 efficacy through Faecalibacterium prausnitzii-derived butyrate and indole-3-propionic acid. | ||
Clin Mol Hepatol Macrophage ATG16L1 expression suppresses MASH progression by promoting lipophagy | ||
Nat Commun Compression-induced NF-κB activation sustains tumor cell survival in confinement by detoxifying aldehydes and promotes metastasis.
| ||
Exp Mol Med PBK drives PARP inhibitor resistance through the TRIM37/NFκB axis in ovarian cancer | ||
Sci Transl Med A peptide immunomodulator activates MST1 to expand and stabilize murine and human regulatory T cells for immune tolerance |
Reviews
What customers say
Both reviewers used this antibody for Western Blot at a 1:1000 dilution and reported positive results. One user (rated 5/5) detected NF-κB p65 in U2OS cells, noting the band appeared at the lowest part of the membrane. The other (rated 4/5) tested it on dendritic cells and highlighted that clear bands were obtained even with low total protein loading (15 µg), indicating good sensitivity. Overall, users found the antibody reliable for WB with strong signal at standard dilution.
The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.
Udesh (Verified Customer) (04-19-2023) | Lowest membrane part has NF-kB p65 in U2OS cells
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Federica (Verified Customer) (01-16-2023) | The band comes out well even when low total protein concentration is loaded (15 ug).
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