Rockland Immunochemicals

antibody

NF-kB p65 Antibody

200-301-065s

25 µL

95.00 USD

monoclonal

mouse

western blot, ELISA, immunohistochemistry

200-301-065S

Anti-NFKB p65 (Rel A) (MOUSE) Monoclonal Antibody - 200-301-065S

NF-kB p65 Antibody

This protein A purified mouse monoclonal antibody is directed against NFkB p65 (Rel A) and a 1:1000 dilution recognizes a 65 kD band by Western blot against HeLa whole cell lysate. Control peptide (100-4165p) is sold separately.

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Monoclonal

27F9.G4

1.0 mg/mL

by UV absorbance at 280 nm

(None)

25 µL

ul

1:50,000-1:100,000

1:200-1:600

1:5000

1:1000 - 1:5000

Expiration date is three (3) months from date of opening.

IgG2a

RELA

Mouse

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.

NFkB p65 (Rel A) peptide corresponding to a region near the C-terminus of the human protein conjugated to Keyhole Limpet Hemocyanin (KLH).

Dry Ice

Liquid (sterile filtered)

0.01% (w/v) Sodium Azide

This product 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.

Human

Store vial at -20° C or below prior to opening. This vial contains a relatively low volume of reagent (25 µL). To minimize loss of volume dilute 1:10 by adding 225 µL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below. Store the vial at -20°C or below after dilution. Avoid cycles of freezing and thawing.

IgG2a kappa

NFKB, nfkb, NF-kB, NF-kappaB, NFkappaB, Nuclear factor NF-kappa-B p65 subunit

Human

NFkappaB was originally identified as a factor that binds to the immunoglobulin kappa light chain enhancer in B cells. It was subsequently found in non-B cells in an inactive cytoplasmic form consisting of NFkappaB bound to IkappaB. NFkappaB was originally identified as a heterodimeric DNA binding protein complex consisting of p65 (RelA) and p50 (NFKB1) subunits. 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. IkappaBalpha-binds to the p65 subunit, preventing nuclear localization and DNA binding. Low levels of p52 and p50 homodimers can also exist in cells.

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.

Peptide

No

Yes

C-Terminus

12/15/2011 7:06 PM

6/24/2015 1:10 PM

ELISA,Immunohistochemistry,IF Microscopy,Western Blot

Primary Antibodies

Unconjugated

95.00

95.00 USD

Q04206

223468676

Anti-NFKB p65 (Rel A) Antibody - Immunofluorescence Microscopy

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.