[KO Validated] NF-kB p65/RelA Rabbit mAb, Unconjugated

Article Name:	[KO Validated] NF-kB p65/RelA Rabbit mAb, Unconjugated
Biozol Catalog Number:	ABB-A22331
Supplier Catalog Number:	A22331
Alternative Catalog Number:	ABB-A22331-100UL,ABB-A22331-20UL,ABB-A22331-500UL,ABB-A22331-1000UL
Manufacturer:	ABclonal
Host:	Rabbit
Category:	Antikörper
Application:	ChIP, ELISA, IF, IP, WB
Species Reactivity:	Human
Immunogen:	Synthetic peptide. This information is considered to be commercially sensitive.
Conjugation:	Unconjugated
Alternative Names:	p65, CMCU, NFKB3, AIF3BL3, lA

NF-kappa-B is a ubiquitous transcription factor involved in several biological processes. It is held in the cytoplasm in an inactive state by specific inhibitors. Upon degradation of the inhibitor, NF-kappa-B moves to the nucleus and activates transcription of specific genes. NF-kappa-B is composed of NFKB1 or NFKB2 bound to either REL, RELA, or RELB. The most abundant form of NF-kappa-B is NFKB1 complexed with the product of this gene, RELA. Four transcript variants encoding different isoforms have been found for this gene.

Clonality:	Monoclonal
Clone Designation:	[ARC51088]
Molecular Weight:	58kDa/59kDa/60kDa
NCBI:	5970
UniProt:	Q04206
Purity:	Affinity purification
Sequence:	LGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISS
Target:	RELA
Antibody Type:	Primary Antibody

Application Dilute:

WB,1:5000 - 1:20000|IF/ICC,1:500 - 1:2000|IP,0.5µg-4µg antibody for 200µg-500µg extracts of whole cells|ChIP,5µg antibody for 10µg-15µg of Chromatin|ELISA,Recommended starting concentration is 1 µg/mL. Please optimize the concentration based on your speci

Application Notes:

Cross-Reactivity: Human,Mouse,Rat,Monkey, ResearchArea: Epigenetics Nuclear Signaling,Transcription Factors,Protein phosphorylation,Cancer,Signal Transduction,Cell Biology Developmental Biology,Apoptosis,Inhibition of Apoptosis,Death Receptor Signaling Pathway,Endocrine Metabolism,Endocrine and metabolic diseases,Obesity,Immunology Inflammation,B Cell Receptor Signaling Pathway,T Cell Receptor Signaling Pathway,Jak-Stat-IL-6 Receptor Signaling Pathway,NF-kB Signaling Pathway,Toll-like Receptor Signaling Pathway,Neuroscience,Neurodegenerative Diseases,Amyloid Plaque and Neurofibrillary Tangle Formation in Alzheimers Disease,Cardiovascular.

	Western blot analysis of lysates from wild type(WT) and NF-kB p65/RelA knockout (KO) HeLa cells, using [KO Validated] NF-kB p65/RelA Rabbit mAb (A22331) at 1:10000 dilution. Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution. Lysates/proteins: 25µg per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit (RM00020). Exposure time: 10s.
	Confocal imaging of PC-12 cells (treated with TNF-alpha) and PC-12 cells (untreated) cells using [KO Validated] NF-kB p65/RelA Rabbit mAb (A22331, dilution 1:2000) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with alpha-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.
	Western blot analysis of various lysates using [KO Validated] NF-kB p65/RelA Rabbit mAb (A22331) at 1:10000 dilution. Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution. Lysates/proteins: 25µg per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit (RM00020). Exposure time: 30s.
	Confocal imaging of HT-1080 cells (treated with TNF-alpha) and HT-1080 cells (untreated) cells using [KO Validated] NF-kB p65/RelA Rabbit mAb (A22331, dilution 1:2000) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with alpha-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.
	Confocal imaging of NIH/3T3 cells (treated with TNF-alpha) and NIH/3T3 cells (untreated) cells using [KO Validated] NF-kB p65/RelA Rabbit mAb (A22331, dilution 1:2000) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with alpha-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.
	Immunoprecipitation of [KO Validated] NF-kB p65/RelA Rabbit mAb from 500 µg extracts of HeLa cells was performed using 2 µg of [KO Validated] NF-kB p65/RelA Rabbit mAb (A22331). Rabbit IgG isotype control (AC042) was used to precipitate the Control IgG sample. IP samples were eluted with 1X Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using [KO Validated] NF-kB p65/RelA Rabbit mAb (A22331) at a dilution of 1:10000.
	Chromatin immunoprecipitation analysis of extracts of HT-1080 cells, HT-1080 cells were treated by TNF-alpha (20 ng/ml) at 37°C for 30 minutes, using [KO Validated] NF-kB p65/RelA Rabbit mAb (A22331) and rabbit IgG.The amount of immunoprecipitated DNA was checked by quantitative PCR. Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.
	Immunoprecipitation of [KO Validated] NF-kB p65/RelA Rabbit mAb from 500 µg extracts of HeLa cells was performed using 2 µg of [KO Validated] NF-kB p65/RelA Rabbit mAb (A22331). Rabbit IgG isotype control (AC042) was used to precipitate the Control IgG sample. IP samples were eluted with 1X Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using [KO Validated] NF-kB p65/RelA Rabbit mAb (A22331) at a dilution of 1:10000.
	Chromatin immunoprecipitation analysis of extracts of HT-1080 cells, HT-1080 cells were treated by TNF-alpha (20 ng/ml) at 37°C for 30 minutes, using [KO Validated] NF-kB p65/RelA Rabbit mAb (A22331) and rabbit IgG.The amount of immunoprecipitated DNA was checked by quantitative PCR. Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.