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Primary Antibodies  >  Transcription Factor Antibodies
 

NF-kB p65 Antibody

Mouse Monoclonal 27F9.G4 IgG2a kappa
Code

200-301-065
Size

100 µg
Price

$309.00
Availability

Ships next business day.
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Rockland Monoclonal anti NFKB p65 (Rel A) antibody was used to detect p65 by immunofluorescence at a dilution of 1:5000.   Hela cells were grown to sub-confluent on 18 mm2 glass coverslips #1.5.  Cells were either unstimulated (A), or stimulated (B) with 50 ng/ml of TNF alpha for 30 min prior fixation.  Cells were then fixed in methanol and blocked with 10% normal goat serum (NGS), in PBS, and TritonX 0.2% (Tx) and incubated for 1 hr at RT with primary ab, counterstained with DAPI and washed in PBS/NGS/Tx.  Cells were incubated for 1 hr at RT with Atto 425 conjugated anti mouse secondary antibody for STED CW imaging.  Data was collected on a STED-CW TCS-SP5 Confocal system equipped with a DFC 350FX camera allowing sequential acquisition in widefiled, confocal and STED CW imaging on the same system.
Rockland anti NFKB p65 (Rel A) monoclonal antibody 200-301-065 was used to detect ~65 kD band (red arrow) in HeLa whole cell lysate.  Lysate was run on 4-20% gradient gel transferred under standard conditions and blocked in 1% BSA-TTBS 30 min RT.  Blot was probed with monoclonal anti p65 1:1000 in 1% BSA-TBS-T o/n 4°C and detected with HRP conjugated Rb-anti Mouse antibody 1:40,000 in MB-070 30 min RT.
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Synonyms: NFKB, nfkb, NF-kB, NF-kappaB, NFkappaB, Nuclear factor NF-kappa-B p65 subunit

NF-kB p65 Antibody Properties

Description
Anti-NFKB p65 (Rel A) (MOUSE) Monoclonal Antibody - 200-301-065
Host
Mouse
Target Species
Human
Known Cross Reactivity
Human
Clonality
Monoclonal 27F9.G4 IgG2a kappa
Application
ELISA : 1:50,000-1:100,000
IF Microscopy : 1:5,000
Western Blot : 1:1,000-1:5,000
Immunochemistry: 1:200-1:600
Other Dilution: User Optimized
Physical State
Liquid (sterile filtered)
Shipping condition
Dry Ice
Format
IgG
Label
Concentration
1.0 mg/mL by UV absorbance at 280 nm
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
0.01% (w/v) Sodium Azide
Stabilizer
None
GeneName
RELA
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Lot#  

NF-kB p65 Antibody Description

Background
NFkappaB was originally identified as a factor that binds to the immunoglobulin kappa light chain enhancer in B cells. Other identified subunits include p52 (NFKB2), c-Rel, and RelB. The p65, cRel, and RelB subunits are responsible for transactivation. The p50 and p52 subunits possess DNA binding activity but limited ability to transactivate. p52 has been reported to form transcriptionally active heterodimers with the NFkappaB subunit p65, similar to p50/p65 heterodimers. The heterodimers of p52/p65 and p50/p65 are regulated by physical inactivation in the cytoplasm by IkappaBalpha. Cell Biology, Nuclear Signaling, Neuroscience and Signal Transduction Research.
Immunogen
NFkB p65 (Rel A) peptide corresponding to a region near the C-terminus of the human protein conjugated to Keyhole Limpet Hemocyanin (KLH).
Immunogen Type
Peptide
Storage Condition
Store NF-kB p65 Antibody at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
Application Note
Anti-NF-kB p65 Antibody is a mouse monoclonal antibody directed against NFkB p65 (Rel A) and recognizes a 65 kD band by Western blot against HeLa whole cell lysate. Control peptide (100-4165p) is sold separately
Purity/Specifity
Anti-NF-kB p65 Antibody was purified from concentrated tissue culture supernate by Protein A chromatography and showed a single band by IEP (immunoelectrophoresis) when tested with anti-mouse antibody. Reactivity was confirmed by ELISA against peptide conjugated carrier protein and by Western blot against HeLa whole cell lysate.
Disclaimer Note-General
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.
General Reference
Unlap, M.T. and Jope, R.S. (1997). Dexamethasone attenuates NF-kappaB DNA binding activity without inducing IkappaB levels in rat brain in vivo. Molecular Brain Research 45: 83-89 Poser, I., Golob, M., Buettner, R. and Bosserhoff, A.K. (2003) Upregulation of HMG1 Leads to Melanoma Inhibitory Activity Expression in Malignant Melanoma Cells and Contributes to Their Malignancy Phenotype. Mol. Cell Biol. 23(8): 2991-2998. Jobin, C, et al. (1999) Curcumin blocks cytokine-mediated NF-kappa B activation and proinflammatory gene expression by inhibiting inhibitory factor I-kappa B kinase activity. J. Immunol. 163(6): 3474-3483.
Specific Reference
Merkhofer EC, Cogswell P, Baldwin AS. Her2 activates NF-kappaB and induces invasion through the canonical pathway involving IKKalpha. Oncogene. 2010 Feb 25;29(8):1238-1248.
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Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, IF, FC
Reacts With
human, mouse, rat, chicken
Catalog Number
100-401-401

Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, GS, Other
Reacts With
human, mouse, rat
Catalog Number
100-4165

Product Type
Primary Antibodies;
Applications
ELISA, WB
Reacts With
human
Catalog Number
100-4165N

Product Type
Proteins and Peptides;
Catalog Number
100-4165NP

Product Label

 

Description
Unconjugated
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry