Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric)

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-29661

Novus Biologicals, part of Bio-Techne
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Key Product Details

Sample Type & Volume Required Per Well

Cell and tissue lysates (0.1 ml)

Sensitivity

15.6 ng/ml

Assay Range

15.6 - 1000 ng/ml

View all RelA/NFkB p65 ELISA Kits »

Product Specifications

Assay Type

Sandwich ELISA

Kit Type

ELISA Kit (Colorimetric)

Reactivity

Human, Mouse, Rat

Specificity

For the detection of cytoplasmic, nuclear and total free NF-kB/p65 in the nucleus of either cells or tissues. Human, mouse, rat and sheep reported in the literature.

Description

APPROPRIATE CONTROLS TO INCLUDE: Following is a list of suggested controls to include with each analysis: 1. No capture antibody added to well. 2. No lysate added to well. 3. No capture antibody or lysate added to well. 4. Positive control: use a cell line or tissue known to constitutively express p65 or a recombinantly expressed p65. 5. Negative control: use a cell line or tissue known to not express p65.

Please note that this kit has not yet been tested or used with any of the reporter cell lines and is therefore not guaranteed for detection of the target protein these samples. Contains reagents and protocol to prepare whole, nuclear, and cytoplasmic cell fractions. Multiple samples can be analyzed in a low-volume, high-throughput format. Full analysis complete in just hours. Allows direct measurement of changes in p65 translocation. Allows study of NF-kB activation without gel-shift assay. [This product is the same as Imgenex's NF-kappaB p65 ActivELISA Kit, IMK-503].

Scientific Data Images for Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric)

ELISA: Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) [NBP2-29661]

ELISA: Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) [NBP2-29661]

ELISA: Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) [NBP2-29661] - Standard curve using the internal RelA standard.
ELISA: Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) [NBP2-29661]

ELISA: Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) [NBP2-29661]

ELISA: Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) [NBP2-29661] - Nuclear translocation of NF-kB p65 with LPS in Jurkat cells. Jurkat cells, grown to a density of 2X10(6) cells/ml, were treated with LPS (15 ug/ml). The cells were harvested at various indicated time points, and cell lysates were tested in Western analysis using NB100-56712 and also in this ELISA Kit.
ELISA: Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) [NBP2-29661]

ELISA: Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) [NBP2-29661]

ELISA: Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) [NBP2-29661] - Immunofluorescence indicates NF-kB p65 subunit was expressed exclusively in AA-G groups and AST-G cytoplasm. High glucose stimulation impact on TSMCs in NF-kB pathway. (A) NF-kB p65 protein subunit expression in TSMCs in each group. (B) NF-kB p65 subunit immunofluorescence. Each number denotes different subunits. Alexa 488 green served as secondary antibodies.Citation: Hara K, Hamada C, Wakabayashi K, Kanda R, Kaneko K, Horikoshi S, et al. (2017) Scavenging of reactive oxygen species by astaxanthin inhibits epithelial-mesenchymal transition in high glucose-stimulated mesothelial cells. PLoS ONE 12(9): e0184332. https://doi.org/10.1371/journal.pone.0184332

Kit Contents for Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric)

  • 1 M DTT (for Nuclear Extraction from tissue) 100 ul
  • 10 mM DTT (for Nuclear Lysis Buffer) 500 ul
  • 10% Detergent Solution 10 ml
  • 100 mM PMSF 10 ml
  • 100X Protease Inhibitor Cocktail (PIC) 100 ul
  • 10X Hypotonic Lysis Buffer 10 ml
  • 10X PBS 2 x 50 ml
  • 1X Nuclear Extraction Buffer 10 ml
  • AKP-Conjugated Secondary Ab 10 ul
  • BSA 2 x 0.5 g
  • Capture Antibody 200ul
  • Coating Buffer 2 x 10 ml
  • Detecting Antibody 200 ul
  • ELISA Plates 2
  • Manual 1
  • pNPP 4 x 5 mg
  • pNPP Substrate Buffer 2 x 10 ml
  • Recombinant p65 Standard 2 vials lyophilized (0.42 ug/vial)

Preparation and Storage

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Storage is content dependent.

Background: RelA/NFkB p65

Activation of the NF-kB pathway can be triggered by many factors including TNFalpha, UV, IL-1, lipopolysaccharide (LPS), mitogens, and phorbol esters. NF-kB is controlled by a family of inhibitory proteins called, IkBs. IkB proteins are phosphorylated by IkB kinase complex consisting of at least three proteins, IKK1/alpha, IKK2/Beta, and IKK3/gamma. External stimuli such as tumor necrosis factor or other cytokines initiate a signal transduction cascade that leads to the activation of IkB-kinase complex, which then specifically phosphorylates IkBalpha on Serine-32 and Serine-36. Phosphorylation of these sites leads to ubiquitination of IkBalpha and subsequent degradation by the 26 S proteasome. Degradation of IkBalpha results in unmasking of the nuclear localization signal of NF-kB dimers, which subsequently translocates to the nucleus and acts as a transcription factor for genes controlling inflammatory cytokines, adhesion molecules, and other proteins. Thus the nuclear levels of p65 may correlate positively with the activation of NF-kB pathway.

Long Name

v-rel Reticuloendotheliosis Viral Oncogene Homolog A

Alternate Names

NFkB p65, NFKB3, p65RelA, nf kb elisa kit, nfkb elisa kit, p65 elisa kit

Gene Symbol

RELA

Additional RelA/NFkB p65 Products

Product Documents for Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric)

Product Datasheets

Certificate of Analysis

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Product Specific Notices for Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. ELISA Kits are guaranteed for 6 months from date of receipt.

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