For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome

Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
United States
Your country/region is currently set to:

If incorrect, please enter your country/region into the box below, to view site information related to your country/region.

Call (888) 77-ABCAM (22226) or contact us
Need help? Contact us

  • My account
  • Sign out
Sign in or Register with us

Welcome

Sign in or

Don't have an account?

Register with us
My basket
Quick order
Abcam homepage

  • Research Products
    By product type
    Primary antibodies
    Secondary antibodies
    ELISA and Matched Antibody Pair Kits
    Cell and tissue imaging tools
    Cellular and biochemical assays
    Proteins and Peptides
    By product type
    Proteomics tools
    Agonists, activators, antagonists and inhibitors
    Cell lines and Lysates
    Multiplex miRNA assays
    Multiplex Assays
    By research area
    Cancer
    Cardiovascular
    Cell Biology
    Epigenetics
    Metabolism
    Developmental Biology
    By research area
    Immunology
    Microbiology
    Neuroscience
    Signal Transduction
    Stem Cells
  • Customized Products & Partnerships
    Customized Products & Partnerships

    Customized products and commercial partnerships to accelerate your diagnostic and therapeutic programs.

    Customized products

    Partner with us

  • Support
    Support hub

    Access advice and support for any research roadblock

    View support hub

    Protocols

    Your experiments laid out step by step

    View protocols

  • Events
    • Conference calendar
    • Cancer
    • Cardiovascular
    • Epigenetics & Nuclear signaling
    • Immunology
    • Neuroscience
    • Stem cells
    • Tradeshows
    • Scientific webinars
    Keep up to date with the latest events

    Full event breakdown with abstracts, speakers, registration and more

    View global event calendar

  • Pathways
    Cell signalling pathways

    View all pathways

    View all interactive pathways

  1. Link

    nf-kappab-p65-ps536--total-elisa-kit-ab176663.pdf

  1. Send me a copy of this email
    I agree to the terms and conditions.
Cell Biology Apoptosis Intracellular NFkB RelA (p65)
Share by email
SimpleStep

NF kappaB p65 (pS536 + Total) ELISA Kit (ab176663)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Submit a question References (13)

Product price, shipping and contact information

Currently unavailable

Sorry, we can't display this right now.

Please contact us to place your order, or try again later.

 

Loading size & price…

 

Shipping and order information

Shipping info

Promotion Information

Abpromise

Guaranteed product quality, expert customer support.

Find out more.

Other - NF kappaB p65 (pS536 + Total) ELISA Kit (ab176663)
  • Typical cell lysate dilution series
  • Linearity of dilution
  • Comparison of NFkB p65  expression in different cell lines
  • NFkB p65 (pS536) phosphorylation in response to TNFa treatment

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 500 pg/ml
  • Sample type: Cell Lysate, Tissue Homogenate
  • Detection method: Colorimetric
  • Assay type: Semi-quantitative
  • Reacts with: Mouse, Human

You may also be interested in

Primary
Product image
Anti-GIV antibody (ab113890)
ELISA
Product image
STAT1 (pS727) + total STAT1 ELISA Kit (ab126455)
ELISA
Product image
BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)

View more associated products

Overview

  • Product name

    NF kappaB p65 (pS536 + Total) ELISA Kit
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    (pS536) 6 3.4%
    (Total) 6 2.8%
    Inter-assay
    Sample n Mean SD CV%
    (pS536) 3 10.3%
    (Total) 3 8.7%
  • Sample type

    Cell Lysate, Tissue Homogenate
  • Assay type

    Semi-quantitative
  • Sensitivity

    500 pg/ml
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Product overview

    Abcam’s NFkB p65 (pS536) and NFkB p65 (Total) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of NFkB p65 (pS536) and Total NFkB p65 protein in Human and mouse cells.


    The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    As of October 2019, this kit was reformulated with new antibodies to maintain continued long term supply.

  • Notes

    Estimated sensitivity: Phospho-NF-κB p65 (Ser536): 5 µg/mL (tested with HeLa), Total NF-κB p65: 5 µg/mL (tested with HeLa)
    Range: Phospho-NF-κB p65 (Ser536):  6-600 µg/mL, Total NF-κB p65: 6-600 µg/mL

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Microplate

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Wash Buffer PT 1 x 15ml
    50X Cell Extraction Enhancer Solution 1 x 1ml
    5X Cell Extraction Buffer PTR 1 x 12ml
    Lyophilized NF kappaB p65 Control Lysate 1 vial
    NF kappaB p65 (pS536) Capture Antibody 1 x 1.5ml
    NF kappaB p65 (pS536) Detector Antibody 1 x 1.5ml
    NF kappaB p65 (Total) Capture Antibody 1 x 1.5ml
    NF kappaB p65 (Total) Detector Antibody 1 x 1.5ml
    Plate Seal 1 unit
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Substrate 1 x 12ml
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • NFkB
    • RelA (p65)
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Other
    • Microbiology
    • Interspecies Interaction
    • Host Virus Interaction
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • NFkB Pathway
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • NFkB pathway
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • NFkB pathway
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Inflammatory mediators
    • Microbiology
    • Interspecies Interaction
    • Host Immune Response
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Signal transduction proteins ELISA kits
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Phosphoprotein ELISA kits
    • Metabolism
    • Types of disease
    • Obesity
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • NFkB
    • RelA (p65)
  • Cellular localization

    NF-kB p65: Nucleus. Cytoplasm. Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor (I-kappa-B). Colocalized with RELA in the nucleus upon TNF-alpha induction. NF-kappaB p65 (Total): Nucleus. Cytoplasm. Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor (I-kappa-B). Colocalized with RELA in the nucleus upon TNF-alpha induction.
  • Database links

    • Entrez Gene: 5970 Human
    • Entrez Gene: 19697 Mouse
    • Entrez Gene: 19697 Mouse
    • Entrez Gene: 309165 Rat
    • Omim: 164014 Human
    • SwissProt: Q04206 Human
    • SwissProt: Q04207 Mouse
    • SwissProt: Q04207 Mouse
    • Unigene: 502875 Human
    • Unigene: 249966 Mouse
    see all

Images

  • Other - NF kappaB p65 (pS536 + Total) ELISA Kit (ab176663)
    Other - NF kappaB p65 (pS536 + Total) ELISA Kit (ab176663)

    SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Typical cell lysate dilution series
    Typical cell lysate dilution series

    Example of a typical NFĸB p65 (pS536) and NFĸB p65 (Total) cell lysate dilution series. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

  • Linearity of dilution
    Linearity of dilution

    Linearity of dilution in representative sample matrices. Cellular lysates were prepared at 3 concentrations in common media containing 1X Cell Extraction Buffer PTR. Data from duplicate measurements of NFĸB p65 (pS536) are normalized and plotted.

  • Comparison of NFkB p65  expression in different cell lines
    Comparison of NFkB p65 expression in different cell lines

    Cell line analysis for Total NFĸB p65 from 200 µg/mL preparations of cell extracts. Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).

  • NFkB p65 (pS536) phosphorylation in response to TNFa treatment
    NFkB p65 (pS536) phosphorylation in response to TNFa treatment

    Induction of NFĸB p65 (pS536) phosphorylation in MCF-7 cells in response to TNFα treatment. HeLa cells were cultured in 96-well tissue culture plates, and treated (10 min) with a dose-range of TNFα before cell lysis. Data from quadruplicate measurements of NFĸB p65 (pS536) are plotted and compared against total NFĸB p65 protein levels. Comparative NFĸB p65 (pS536) and NFĸB p65 (Total) data also shown by Western Blot.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (13)

Publishing research using ab176663? Please let us know so that we can cite the reference in this datasheet.

ab176663 has been referenced in 13 publications.

  • Ting HK  et al. Inflammatory Regulation by TNF-a-Activated Adipose-Derived Stem Cells in the Human Bladder Cancer Microenvironment. Int J Mol Sci 22:N/A (2021). PubMed: 33924332
  • Li H  et al. Curcumin plays a protective role against septic acute kidney injury by regulating the TLR9 signaling pathway. Transl Androl Urol 10:2103-2112 (2021). PubMed: 34159091
  • Yang L  et al. Perampanel, an AMPAR antagonist, alleviates experimental intracerebral hemorrhage-induced brain injury via necroptosis and neuroinflammation. Mol Med Rep 24:N/A (2021). PubMed: 34080030
  • Feng X  et al. Dexmedetomidine alleviates early brain injury following traumatic brain injury by inhibiting autophagy and neuroinflammation through the ROS/Nrf2 signaling pathway. Mol Med Rep 24:N/A (2021). PubMed: 34278508
  • Saber S  et al. The Supportive Role of NSC328382, a P2X7R Antagonist, in Enhancing the Inhibitory Effect of CRID3 on NLRP3 Inflammasome Activation in Rats with Dextran Sodium Sulfate-Induced Colitis. J Inflamm Res 14:3443-3463 (2021). PubMed: 34321905
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

There are currently no Customer reviews or Questions for ab176663.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Get resources and offers direct to your inbox Sign up
A-Z by research area
  • Cancer
  • Cardiovascular
  • Cell biology
  • Developmental biology
  • Epigenetics & Nuclear signaling
  • Immunology
  • Metabolism
  • Microbiology
  • Neuroscience
  • Signal transduction
  • Stem cells
A-Z by product type
  • Primary antibodies
  • Secondary antibodies
  • Biochemicals
  • Isotype controls
  • Flow cytometry multi-color selector
  • Kits
  • Loading controls
  • Lysates
  • Peptides
  • Proteins
  • Slides
  • Tags and cell markers
  • Tools & Reagents
Help & support
  • Support
  • Make an Inquiry
  • Protocols & troubleshooting
  • Placing an order
  • RabMAb products
  • Biochemical product FAQs
  • Training
  • Browse by Target
Company
  • Corporate site
  • Investor relations
  • Company news
  • Careers
  • About us
  • Blog
Events
  • Tradeshows
  • Conferences
International websites
  • abcam.cn
  • abcam.co.jp

Join with us

  • LinkedIn
  • facebook
  • Twitter
  • YouTube
  • Terms of sale
  • Website terms of use
  • Cookie policy
  • Privacy policy
  • Legal
  • Modern slavery statement
© 1998-2022 Abcam plc. All rights reserved.