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NF-kB p65 antibody (AA 136-224)

NFkBP65 Reactivity: Human, Rat, Dog, Rabbit, Frog WB, BI Host: Mouse Monoclonal 20-NF unconjugated
Catalog No. ABIN968223
  • Target See all NF-kB p65 (NFkBP65) Antibodies
    NF-kB p65 (NFkBP65) (Nuclear Factor-kB p65 (NFkBP65))
    Binding Specificity
    • 40
    • 31
    • 29
    • 28
    • 28
    • 25
    • 23
    • 23
    • 20
    • 17
    • 16
    • 15
    • 14
    • 11
    • 9
    • 9
    • 8
    • 8
    • 5
    • 5
    • 5
    • 4
    • 4
    • 4
    • 4
    • 3
    • 3
    • 3
    • 3
    • 3
    • 3
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    AA 136-224
    Reactivity
    • 531
    • 371
    • 297
    • 41
    • 38
    • 29
    • 24
    • 20
    • 18
    • 15
    • 14
    • 11
    • 5
    • 2
    • 1
    • 1
    Human, Rat, Dog, Rabbit, Frog
    Host
    • 492
    • 43
    • 4
    • 2
    • 1
    Mouse
    Clonality
    • 458
    • 83
    Monoclonal
    Conjugate
    • 292
    • 30
    • 23
    • 18
    • 17
    • 17
    • 17
    • 17
    • 17
    • 17
    • 14
    • 10
    • 10
    • 10
    • 10
    • 10
    • 9
    • 2
    • 2
    This NF-kB p65 antibody is un-conjugated
    Application
    • 425
    • 203
    • 135
    • 118
    • 110
    • 104
    • 92
    • 78
    • 76
    • 29
    • 25
    • 9
    • 7
    • 7
    • 7
    • 6
    • 4
    • 3
    • 2
    • 1
    • 1
    Western Blotting (WB), BioImaging (BI)
    Cross-Reactivity
    Dog (Canine), Frog, Rabbit, Rat (Rattus)
    Characteristics
    1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
    2. Please refer to us for technical protocols.
    3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
    4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
    5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
    6. Triton is a trademark of the Dow Chemical Company.
    Purification
    The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
    Immunogen
    Human NF-kappaB aa. 136-224
    Clone
    20-NF
    Isotype
    IgG1
    Top Product
    Discover our top product NFkBP65 Primary Antibody
  • Application Notes
    Bioimaging
    1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
    2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
    3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
    4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
    5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
    6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
    7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
    8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
    9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
    10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
    11. View and analyze the cells on an appropriate imaging instrument.
    Comment

    Related Products: ABIN968537, ABIN967389

    Restrictions
    For Research Use only
  • Format
    Liquid
    Concentration
    250 μg/mL
    Buffer
    Aqueous buffered solution containing BSA, glycerol, and ≤0.09 % sodium azide.
    Preservative
    Sodium azide
    Precaution of Use
    This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    Storage
    -20 °C
    Storage Comment
    Store undiluted at -20°C.
  • Nishibe, Parry, Ishida, Aziz, Murray, Patel, Rahman, Strand, Saito, Saito, Hammond, Savidge, Mackman, Wijelath: "Oncostatin M promotes biphasic tissue factor expression in smooth muscle cells: evidence for Erk-1/2 activation." in: Blood, Vol. 97, Issue 3, pp. 692-9, (2001) (PubMed).

    Pimentel-Muiños, Seed: "Regulated commitment of TNF receptor signaling: a molecular switch for death or activation." in: Immunity, Vol. 11, Issue 6, pp. 783-93, (2000) (PubMed).

    Kieran, Blank, Logeat, Vandekerckhove, Lottspeich, Le Bail, Urban, Kourilsky, Baeuerle, Israël: "The DNA binding subunit of NF-kappa B is identical to factor KBF1 and homologous to the rel oncogene product." in: Cell, Vol. 62, Issue 5, pp. 1007-18, (1990) (PubMed).

    Shirakawa, Mizel: "In vitro activation and nuclear translocation of NF-kappa B catalyzed by cyclic AMP-dependent protein kinase and protein kinase C." in: Molecular and cellular biology, Vol. 9, Issue 6, pp. 2424-30, (1989) (PubMed).

    Baeuerle, Baltimore: "Activation of DNA-binding activity in an apparently cytoplasmic precursor of the NF-kappa B transcription factor." in: Cell, Vol. 53, Issue 2, pp. 211-7, (1988) (PubMed).

  • Target
    NF-kB p65 (NFkBP65) (Nuclear Factor-kB p65 (NFkBP65))
    Alternative Name
    NF-kappa B p65 (NFkBP65 Products)
    Synonyms
    C-Rel antibody, NFKB3 antibody, p65 antibody, NFkB antibody, c-rel antibody, zgc:100833 antibody, Xrel2 antibody, Xrel3 antibody, rel antibody, rel-A antibody, rel2 antibody, rel3 antibody, v-rel antibody, xrel antibody, REL proto-oncogene, NF-kB subunit antibody, RELA proto-oncogene, NF-kB subunit antibody, v-rel reticuloendotheliosis viral oncogene homolog A (avian) antibody, v-rel avian reticuloendotheliosis viral oncogene homolog antibody, v-rel avian reticuloendotheliosis viral oncogene homolog L homeolog antibody, REL antibody, RELA antibody, Rela antibody, Rel antibody, rel antibody, rel.L antibody
    Background
    NF-kappaB is a ubiquitously expressed transcription factor that regulates many cytokine and Ig genes. It is involved in immune, inflammatory, viral, and acute phase responses. The most studied NF-kappaB complex consists of the p50 and p65 subunits, both containing a 300 amino acid region with homology to the Rel proto-oncogene product. The p50 subunit binds DNA, whereas the p65 subunit is responsible for the interaction of NF-kappaB with its inhibitor, IkappaB. In most cell types, the p50/p65 heterodimer is located within the cytoplasm complexed to IkappaB. This complex prevents nuclear translocation and activity of NF-kappaB. In response to stimuli such as cytokines, LPS, and viral infections, IkappaB is phosphorylated at critical residues. This phosphorylation induces dissociation of the IkappaB/NF-kappaB complex, allowing the free heterodimeric NF-kappaB to form a heterotetramer that translocates to the nucleus. In the nucleus, it binds to the kappaB site within promoters and enhancers and functions as a transcriptional activator.
    Molecular Weight
    65 kDa
    Pathways
    NF-kappaB Signaling, RTK Signaling, TCR Signaling, TLR Signaling, Fc-epsilon Receptor Signaling Pathway, Neurotrophin Signaling Pathway, Activation of Innate immune Response, Cellular Response to Molecule of Bacterial Origin, Hepatitis C, Toll-Like Receptors Cascades, S100 Proteins
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