GeneTex
United States (US)

NFkB p65 antibody

KO/KD Overexpression IP/MS
GTX107678_39981_IP_19011506_773.jpg
GTX107678_43068_20180126_WB_competitor_watermark_19011506_620.jpg
GTX107678_IHC_19011506_884.jpg
GTX107678_43068_20180608_WB_M_19011506_542.jpg
GTX107678_39981_IFA_19011506_388.jpg
GTX107678_43005_20171103_WB_19011506_481.jpg
GTX107678_39981_20150410_IFA_19011506_946.jpg
GTX107678_43068_20180608_WB_KO_watermark_19011506_556.jpg
GTX107678_43068_20180608_WB_R_19011507_154.jpg
GTX107678_41556_WB_B_19011507_398.jpg

Cat No. GTX107678

Host Rabbit
Clonality Polyclonal
Isotype IgG
Application WB, ICC/IF, IHC-P, IP, EMSA
Reactivity Human, Mouse, Rat
APPLICATION

Application Note

*Optimal dilutions/concentrations should be determined by the researcher.
Application Dilution
WB 1:500-1:3000
ICC/IF 1:100-1:1000
IHC-P 1:100-1:1000
IP 1:500-1:1000
EMSA Assay dependent
Not tested in other applications.

Calculated MW

60 kDa. ( Note )

Positive Control

293T , A431 , HeLa , HepG2 , Neuro2A , NIH-3T3 , Raw264.7 , C2C12 , PC-12 , Rat2 , GFP-human NF-kappa-B-transfected 293T

Specificity/Sensitivity

IP/MS validation was supported by references (PMID:30377388)

Predict Reactivity

Bovine, Pig, Chimpanzee(>80% identity)
PROPERTIES

Form

Liquid

Buffer

1XPBS, 20% Glycerol (pH7). 0.025% ProClin 300 was added as a preservative.

Storage

Store as concentrated solution. Centrifuge briefly prior to opening vial. For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, aliquot and store at -20ºC or below. Avoid multiple freeze-thaw cycles.

Concentration

0.86mg/ml(Please refer to the vial label for the specific concentration.)

Antigen Species

Human

Immunogen

Recombinant protein encompassing a sequence within the center region of human NFkB p65. The exact sequence is proprietary.

Purification

Purified by antigen-affinity chromatography.

Conjugation

Unconjugated

Note

For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
TARGET

Synonyms

MGC131774 antibody, NFKB3 antibody, p65 antibody, RELA antibody, nuclear factor NF-kappa-B p65 subunit antibody, transcription factor p65 antibody, NF-kappa-B p65delta3 antibody, nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 antibody, v-rel reticuloendotheliosis viral oncogene homolog A (avian) antibody

Cellular Localization

Nucleus , Cytoplasm

Background

NFKB1 (MIM 164011) or NFKB2 (MIM 164012) is bound to REL (MIM 164910), RELA, or RELB (MIM 604758) to form the NFKB complex. The p50 (NFKB1)/p65 (RELA) heterodimer is the most abundant form of NFKB. The NFKB complex is inhibited by I-kappa-B proteins (NFKBIA, MIM 164008 or NFKBIB, MIM 604495), which inactivate NFKB by trapping it in the cytoplasm. Phosphorylation of serine residues on the I-kappa-B proteins by kinases (IKBKA, MIM 600664, or IKBKB, MIM 603258) marks them for destruction via the ubiquitination pathway, thereby allowing activation of the NFKB complex. Activated NFKB complex translocates into the nucleus and binds DNA at kappa-B-binding motifs such as 5-prime GGGRNNYYCC 3-prime or 5-prime HGGARNYYCC 3-prime (where H is A, C, or T; R is an A or G purine; and Y is a C or T pyrimidine).[supplied by OMIM]

Database

Research Area

DATA IMAGES
GTX107678_39981_IP_19011506_773.jpg

GTX107678 IP Image

NFkB p65 antibody immunoprecipitates NFkB p65 protein in IP experiments. IP Sample: 1000 µg HeLa whole cell lysate/extract A. 40 μg HeLa whole cell lysate/extract B. Control with 2.5 μg of preimmune rabbit IgG C. Immunoprecipitation of NFkB p65 protein by 2.5 μg of NFkB p65 antibody (GTX107678) 10% SDS-PAGE The immunoprecipitated NFkB p65 protein was detected by NFkB p65 antibody (GTX107678) diluted at 1:1000. EasyBlot anti-rabbit IgG (GTX221666-01) was used as a secondary reagent.

GTX107678_43068_20180126_WB_competitor_watermark_19011506_620.jpg

GTX107678 WB Image

Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membranes were blotted with NFkB p65 antibody (GTX107678) diluted at 1:1000 and competitor's antibody (#4764) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

*The competitor is not affiliated with GeneTex and does not endorse this product.

GTX107678_IHC_19011506_884.jpg

GTX107678 IHC-P Image

Immunohistochemical analysis of paraffin-embedded SAS xenograft, using NFkB p65 (GTX107678) antibody at 1:500 dilution.
Antigen Retrieval: Trilogy™ (EDTA based, pH 8.0) buffer, 15min

GTX107678_43068_20180608_WB_M_19011506_542.jpg

GTX107678 WB Image

Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with NFkB p65 antibody (GTX107678) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX107678_39981_IFA_19011506_388.jpg

GTX107678 ICC/IF Image

Confocal immunofluorescence analysis (Olympus FV10i) of paraformaldehyde-fixed U2OS, using NFkB p65(GTX107678) antibody (Green) at 1:500 dilution. Alpha-tubulin filaments were labeled with GTX11304 (Red) at 1:2000.

GTX107678_43005_20171103_WB_19011506_481.jpg

GTX107678 WB Image

Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with NFkB p65 antibody (GTX107678) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX107678_39981_20150410_IFA_19011506_946.jpg

GTX107678 ICC/IF Image

NFkB p65 antibody detects NFkB p65 protein at cytoplasm and nucleus by immunofluorescent analysis.
Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min.
Green: NFkB p65 protein stained by NFkB p65 antibody (GTX107678) diluted at 1:1000.
Red: phalloidin, a cytoskeleton marker, stained by phalloidin (invitrogen, A12380) diluted at 1:200.
Blue: Hoechst 33342 staining.

GTX107678_43068_20180608_WB_KO_watermark_19011506_556.jpg

GTX107678 WB Image

KO/KD

Wild-type (WT) and NFkB p65 knockout (KO) HeLa cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with NFkB p65 antibody (GTX107678) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.
CRISPR knockout cell lines generated by www.edigene.com

GTX107678_43068_20180608_WB_R_19011507_154.jpg

GTX107678 WB Image

Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with NFkB p65 antibody (GTX107678) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX107678_41556_WB_B_19011507_398.jpg

GTX107678 WB Image

Overexpression

NFkB p65 antibody detects NFkB p65 protein by western blot analysis. Non-transfected (-) and NFkB p65 -transfected (+, including GFP-tag) 293T whole cell extracts (15 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with NFkB p65 antibody (GTX107678) diluted by 1:5000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

REFERENCE
REVIEW

NFkB p65 antibody Cat No. GTX107678

Rating ( Average 4.5 based on 2 users reviews)
Application
Western Blot(WB) ( Average 4.5 based on 2 users reviews)
Date : Anonymous submitted on 08-Jan-2018
Score :
Application Tested : WB
Sample Species : Hu
Sample : PANC1 cells
Amount used : 25 μg
Blocking : 5% milk in TBST, 25°C, 1Hr
Primary Antibody : 1:500, 4°C, overnightHr
Date : Anonymous submitted on 15-Mar-2017
Score :
Application Tested : WB
Sample Species : Hu
Sample : SAS cells
Amount used : 15 μg
Blocking : 3% BSA in TBST, 25°C, 1Hr
Primary Antibody : 1:500, 4°C, overnightHr
Application Reference
SDS
PBS.pdf
Glycerol.pdf
Proclin.pdf
Package List Price ($)
$ 319
$ 169