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Purified anti-NF-κB p65 Antibody

Pricing & Availability
Clone
Poly6226 (See other available formats)
Regulatory Status
RUO
Other Names
NFκB p65, nuclear factor kappa light chain enhancer in B cells p65
Isotype
Rabbit Polyclonal IgG
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Product Citations
publications
1_poly-p65_101807
Hela cells stained with purified rabbit polyclonal antibody against NF-κB p65 (poly6226), followed by Rhodamine Red-X conjugated goat anti-rabbit IgG and DAPI
  • 1_poly-p65_101807
    Hela cells stained with purified rabbit polyclonal antibody against NF-κB p65 (poly6226), followed by Rhodamine Red-X conjugated goat anti-rabbit IgG and DAPI
  • 2_NFkB_p65_060204
    Total lysates (15 µg protein) from HeLa (lane 1), Raw 264.7 (lane 2) and UMR106 cells (lane 3) were resolved by electrophoresis (4-20% Tris-Glycine gel), transferred to nitrocellulose, and probed with 0.05 µg/mL Purified anti-NF-κB p65 Antibody, clone poly6226 (upper). Proteins were visualized by chemiluminescence detection using a 1:3000 diluted goat anti-rabbit-IgG secondary antibody conjugated to HRP for the anti-NF-κB p65 Antibody. 1:5000 diluted Direct-Blot HRP anti-β-Actin Antibody (2F1-1) was used as a loading control (lower). Lane M: Molecular weight ladder.
  • 3_Poly6226_GoChIP_NF-kB_Antibody_1_020118.png
    Chromatin Immunoprecipitation (ChIP) was performed using Go-ChIP-Grade™ protein G Enzymatic kit by loading 3 µg of cross-linked chromatin samples from HeLa cells treated with TNFα with either A) 1:100 dilution of Go-ChIP-Grade™ Purified anti-NF-κB p65 (Clone Poly6226), or B) equal amount of Purified Rabbit Polyclonal Isotype Control Antibody (Cat. No. 910801). The enriched DNA was purified and quantified by real-time qPCR using primers targeting human IKBA gene region. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the 5% of total amount of input chromatin.
Cat # Size Price Quantity Check Availability Save
622601 50 µL $118
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622602 200 µL $293
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Description

NF-κB/p65 (nuclear factor kappa light chain enhancer in B cells p65) is a member of the Rel/dorsal family. This ubiquitously expressed nuclear protein is one subunit of the NF-kappa B complex consisting of a 65 kD transactivating subunit and a 50 kD DNA binding subunit. Both subunits are derived from larger precursor proteins. NF-κB acts as a transcriptional activator and was originally identified as an activator of kappa light chain in B cells. NF-κB/p65 is bound (with p50) to IκB inhibitor in cytoplasm in inactive form. Phosphorylation of IκB releases and NF-κB which goes to the nucleus to activate gene expression. NF-κB can be activated with LPS, TNF-α, phorbol ester, or IL-1. The Poly6226 antibody reacts with the C-terminal region of human, mouse, and rat NF-κB p65 and has been shown to be useful for Western blotting.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Mouse, Rat, Human
Antibody Type
Polyclonal
Host Species
Rabbit
Immunogen
peptide-KLH, C-terminal
Formulation
This antibody is provided in phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and 0.2% gelatin.

Previous lots of this product may have been formulated with 0.1% or 0.05% NaN3 instead of 0.09% NaN3. For further information please contact BioLegend Technical Support or Customer Service.
Preparation
The antibody was purified by antigen-affinity chromatography.
Concentration
Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
Storage & Handling
Upon receipt, store undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC, ChIP - Verified
IHC-F, IP - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.05 µg per mL. For immunocytochemistry, the suggested use of the reagent is a dilution range of 1:100~1:400. For ChIP applications, the suggested dilution is 1:50-1:200 by volume. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This product may contain other non-IgG subtypes.

Additional Product Notes

The 50 µL size can be used for approximately 5 Western blots, and the 200 µL size can be used for approximately 20 Western blots.

Application References

(PubMed link indicates BioLegend citation)
  1. Chen R, et al. 2014. J Mol Cell Cardiol. 72:263. PubMed
  2. Tan GK, et al. 2014. Acta Biomater. 10:2684. PubMed
  3. Abecassis A, et al. 2014. Cell Mol Immunol. 4:377. PubMed
  4. Sumedha N, et al. 2014. Hum Exp Toxicol. 34:506. PubMed
  5. Hua X, et al. 2015. PloS One. 10:128039. PubMed
Product Citations
  1. Fang SY, et al. 2021. Exp Ther Med. 369:21. PubMed
  2. Smaldone G, et al. 2021. Sci Rep. 11:18237. PubMed
  3. Haddad H, et al. 2021. Int J Mol Sci. 22:. PubMed
  4. Hua X, et al. 2015. PLoS One. 10:128039. PubMed
  5. Abecassis A, et al. 2014. Cell Mol Immunol. 4:377. PubMed
  6. Chung H, et al. 2021. Nat Methods. 18:1204. PubMed
  7. Afzal SM, et al. 2020. Environ Toxicol. :. PubMed
  8. Shaker ME, et al. 2022. Saudi Pharm J. 30:291. PubMed
  9. Chaudhari S, et al. 2020. Am J Physiol Renal Physiol. 318:F1478. PubMed
  10. Ayari C, et al. 2011. J Urol. 185:1915. PubMed
  11. Ge Y, et al. 2013. J Virol. 87:11160. PubMed
  12. Miltonprabu N 2014. Hum Exp Toxicol. 34:506. PubMed
  13. Yang Z, et al. 2016. Sci Rep. 6:30263. PubMed
  14. Muri J, et al. 2020. eLife. 9:e53627.. PubMed
  15. Tortola L, et al. 2022. Mucosal Immunol. 15:656. PubMed
  16. Tan G, Tabata Y 2014. Acta Biomater. 10:2684. PubMed
  17. Chen R, et al. 2014. J Mol Cell Cardiol. 72:263. PubMed
RRID
SCR_001134 (BioLegend Cat. No. 622601)
SCR_001134 (BioLegend Cat. No. 622602)

Antigen Details

Structure
Member of the Rel/dorsal family. One subunit of the NF-kappa B complex consisting of a 65 kD transactivating subunit and a 50 kD DNA binding subunit. Both subunits derived from larger precursor proteins
Distribution

Ubiquitously expressed nuclear protein

Function
Transcriptional activator, originally identified as an activator of kappa light chain in B cells
Interaction
Interacts with NF-κB p50 and IκB
Biology Area
Cell Biology, Neuroscience, Neuroscience Cell Markers, Transcription Factors
Molecular Family
Nuclear Markers
Antigen References

1. Baldwin AS, et al. 1996. Annu. Rev. Immunol. 14:649 (review).
2. Chen F, et al. 1999. Clin. Chem. 45:7 (review).

Regulation
Bound (with p50) to IkB inhibitor in cytoplasm in inactive form. Phosphorylation of IkB releases and NF-kB goes to the nucleus to activate gene expression. Stimulation with LPS, TNF-α, phorbol ester, or IL-1 activates
Gene ID
4790 View all products for this Gene ID
UniProt
View information about NF-kappaB p65 on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 8    Revision Date: 02/08/2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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