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NF?B p65 phospho Ser536 antibody (pAb)

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for NFkB p65 phospho Ser536 antibody (pAb)Application Notes


Validated Applications:
WB: 1:500 - 1:1,000 dilution

for NFkB p65 phospho Ser536 antibody (pAb)Immunogen

This NFkB p65 phospho Ser536 antibody was raised against a peptide containing phospho-serine 536 of human NFkB p65.

for NFkB p65 phospho Ser536 antibody (pAb)Buffer

Rabbit serum containing 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

for NFkB p65 phospho Ser536 antibody (pAb)References

 
NF?B p65 phospho Ser536 antibody (pAb) tested by Western blot.

NF?B p65 phospho Ser536 pAb tested by Western blot.
HeLa cell extract (20 µg per lane) probed with either NF?B p65 phospho Ser536 pAb (Lanes 1 and 2, 1:500 dilution) or with NF?B p65 pAb (Catalog No. 39369, Lanes 3 and 4, 1:5,000 dilution). Lanes 1 and 3: extracts derived from untreated cells. Lanes 2 and 4: extracts derived from cells treated with TNF-? and Calyculin.

NF?B p65 phospho Ser536 antibody (pAb) tested by dot blot analysis.

NF?B p65 phospho Ser536 pAb tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of of NF?B p65 phospho Ser536 pAb for NF?B p65 phosphorylated at serine 536. Phosphorylated peptides corresponding to the immunogen and related peptides were spotted onto PVDF and probed with NF?B p65 phospho Ser536 pAb at 1:10,000. The amount of peptide (picomoles) spotted is indicated next to each row.

Lane 1: phospho Ser276 NF?B p65 peptide.
Lane 2: unmodified peptide surrounding Ser276 NF?B p65 peptide.
Lane 3: phospho Ser529 NF?B p65 peptide.
Lane 4: unmodified peptide surrounding Ser529 NF?B p65.
Lane 5: phospho Ser536 NF?B p65 peptide.
Lane 6: unmodified peptide surrounding Ser536 NF?B p65.
Lane 7: phospho Ser337 NF?B p50 peptide.
Lane 8: unmodified peptide surrounding Ser337 NF?B p50.

for NFkB p65 phospho Ser536 antibody (pAb)Background

NF?B p65 (Nuclear factor NF-kappa-B p65 subunit also known as NF?B p65 or RelA) is a member of the Rel family of transcription factors that are involved in processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. The Rel family includes c-Rel, RelA (NF?B p65, NFkB3) and RelB (I-Rel). Rel proteins form dimers with the NF?B p50 (proteolytically processed from p105/NF?B1)and p52 (proteolytically processed from p100/NF?B2) members of the NF?B family that bind DNA and activate transcription. The NF?B p65/p50 heterodimer is the most abundant form of NF?B.

NF?B signaling is negatively regulated by the sequestration of the NF?B complex in the cytoplasm by its association with the I?B family of inhibitory proteins. The I?B Kinase (IKK) complex is the key enzyme involved in the activation and translocation of NF?B. The IKK complex is composed of two catalytic subunits (IKK? and IKK?) and a regulatory subunit (IKK?). Upon its activation, the IKK complex phosphorylates I?B proteins (I?B?, I?B? phospho Ser32, 36), which marks them for ubiquitination and degradation. This enables the NF?B complex to translocate to the nucleus where it regulates transcription by binding to DNA.

Phosphorylation at serine 536 stimulates acetylation at lysine 310 and interaction with the acetyltransferase CBP. The phosphorylated and acetylated forms show enhanced transcriptional activity.

for NFkB p65 phospho Ser536 antibody (pAb)Positive Control

Active Motif's HeLa nuclear extract (TNF-? stimulated) (Catalog No. 40210) can be used as a positive control for NF?B p65 phospho Ser536 antibody.

for NFkB p65 phospho Ser536 antibody (pAb)Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

for NFkB p65 phospho Ser536 antibody (pAb)Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting