- 33-9900 - Invitrogen Antibodies RELA antibody

Submitted references Estrogen Receptor Beta Influences the Inflammatory p65 Cistrome in Colon Cancer Cells.

Indukuri R, Hases L, Archer A, Williams C
Frontiers in endocrinology 2021;12:650625

NF-κB activation in retinal microglia is involved in the inflammatory and neovascularization signaling in laser-induced choroidal neovascularization in mice.

Hikage F, Lennikov A, Mukwaya A, Lachota M, Ida Y, Utheim TP, Chen DF, Huang H, Ohguro H
Experimental cell research 2021 Jun 1;403(1):112581

Cyclin F Downregulation Affects Epithelial-Mesenchymal Transition Increasing Proliferation and Migration of the A-375 Melanoma Cell Line.

Krajewski A, Gagat M, Mikołajczyk K, Izdebska M, Żuryń A, Grzanka A
Cancer management and research 2020;12:13085-13097

Epigenetic Suppression of HIV in Myeloid Cells by the BRD4-Selective Small Molecule Modulator ZL0580.

Alamer E, Zhong C, Liu Z, Niu Q, Long F, Guo L, Gelman BB, Soong L, Zhou J, Hu H
Journal of virology 2020 May 18;94(11)

Partitioning of MLX-Family Transcription Factors to Lipid Droplets Regulates Metabolic Gene Expression.

Mejhert N, Kuruvilla L, Gabriel KR, Elliott SD, Guie MA, Wang H, Lai ZW, Lane EA, Christiano R, Danial NN, Farese RV Jr, Walther TC
Molecular cell 2020 Mar 19;77(6):1251-1264.e9

Structure-guided drug design identifies a BRD4-selective small molecule that suppresses HIV.

Niu Q, Liu Z, Alamer E, Fan X, Chen H, Endsley J, Gelman BB, Tian B, Kim JH, Michael NL, Robb ML, Ananworanich J, Zhou J, Hu H
The Journal of clinical investigation 2019 Jul 22;129(8):3361-3373

A Single Nucleotide Polymorphism in the Vitamin D Receptor Gene Is Associated With Decreased Levels of the Protein and a Penetrating Pattern in Crohn's Disease.

Gisbert-Ferrándiz L, Salvador P, Ortiz-Masiá D, Macías-Ceja DC, Orden S, Esplugues JV, Calatayud S, Hinojosa J, Barrachina MD, Hernández C
Inflammatory bowel diseases 2018 Jun 8;24(7):1462-1470

Role of p62/SQSTM1 beyond autophagy: a lesson learned from drug-induced toxicity in vitro.

Alegre F, Moragrega ÁB, Polo M, Marti-Rodrigo A, Esplugues JV, Blas-Garcia A, Apostolova N
British journal of pharmacology 2018 Feb;175(3):440-455

Glycocalyx Degradation Induces a Proinflammatory Phenotype and Increased Leukocyte Adhesion in Cultured Endothelial Cells under Flow.

McDonald KK, Cooper S, Danielzak L, Leask RL
PloS one 2016;11(12):e0167576

Antipsychotic drugs suppress the AKT/NF-κB pathway and regulate the differentiation of T-cell subsets.

Chen ML, Tsai TC, Lin YY, Tsai YM, Wang LK, Lee MC, Tsai FM
Immunology letters 2011 Oct 30;140(1-2):81-91

Cautionary notes on the use of NF-κB p65 and p50 antibodies for CNS studies.

Herkenham M, Rathore P, Brown P, Listwak SJ
Journal of neuroinflammation 2011 Oct 14;8:141

Sequence characteristics of functional siRNAs.

Jagla B, Aulner N, Kelly PD, Song D, Volchuk A, Zatorski A, Shum D, Mayer T, De Angelis DA, Ouerfelli O, Rutishauser U, Rothman JE
RNA (New York, N.Y.) 2005 Jun;11(6):864-72

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of A-431 (Lane 1), U-87 MG (Lane 2), A549 (Lane 3), Raji (Lane 4), K-562 (Lane 5) and NIH/3T3 (Lane 6). The blot was probed with Anti-NFkB p65 Monoclonal Antibody (Product # 33-9900, 1 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 65 kDa band corresponding to NFkB p65 was observed across the cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of NF-kappa-B (p65) was performed by loading 20 µg of HEK-293 (lane1), A549 (lane2), Jurkat (lane3), HeLa (lane4), NIH/3T3 (lane5) and Raji (lane6) cell lysates using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. NF-kappa-B (p65) was detected at ~65 kDa using NF-kappa-B (p65) Mouse Monoclonal Antibody (Product # 33-9900) at 0.5-1 µg/mL in 2.5 % skim milk at 4°C overnight on a rocking platform. Goat Anti-Mouse IgG - HRP Secondary Antibody (Product # 62-6520) at 1:4000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of NFkB p65 (Fig. a) was performed by loading 30 µg of HeLa control (lane 1), HeLa NFkB p65 knockout (lane 2) whole cell extracts. NFkB p65 was detected at 65 kDa using NFkB p65 mouse monoclonal antibody (Product # 33-9900, 1 µg/mL) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody HRP conjugate (Product # A28177, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Loss of signal in CRISPR mediated knockout (KO) confirms that antibody is specific to NFkB p65.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of A-431 (Lane 1), U-87 MG (Lane 2), A549 (Lane 3), Raji (Lane 4), K-562 (Lane 5) and NIH/3T3 (Lane 6). The blot was probed with Anti-NFkB p65 Monoclonal Antibody (Product # 33-9900, 1 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 65 kDa band corresponding to NFkB p65 was observed across the cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of NFkB p65 was achieved by transfecting HeLa with NFkB p65 specific siRNAs (Silencer® select Product # s11916, s11914). Western blot analysis was performed using Nuclear enriched extracts from the NFkB p65 knockdown cells (lane 3), scrambled siRNA transfected cells (lane 2), and untransfected cells (lane 1). The blot was probed with NFkB p65 Monoclonal Antibody (2A12A7) (Product # 33-9900, 0.5 µg/mL) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:20000 dilution). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to NFkB p65.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of NF-kappa-B (p65) showing staining in the Cytoplasm and Nucleus of paraffin-embedded human breast carcinoma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a NF-kappa-B (p65) mouse monoclonal antibody (Product # 33-9900) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of NF-kappa-B (p65) showing staining in the Cytoplasm and Nucleus of paraffin-embedded mouse spleen tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a NF-kappa-B (p65) mouse monoclonal antibody (Product # 33-9900) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: ChIP- qPCR analysis of NF-kappa-B-p65 was performed with 3 µg/mL of the NF-kappa-B-p65 mouse monoclonal Antibody (Product # 33-9900) on sheared chromatin from 2 million HeLa cells treated with TNF-a (50 ng/mL for 1h) using the MAGnify Chromatin Immunoprecipitation System (Product # 49-2024). Normal mouse IgG (3 µg/mL) was used as a negative IP control. The purified DNA from each ChIP sample was analyzed by StepOnePlus Real-Time PCR System (Product # 4376600) with primers for the promoter of active IL-8 and IL-6 gene, used as positive control targets, and the GAPDH gene, used as negative control target. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Genomic distribution of p65 binding sites in colon cell lines. (A) Identified p65 binding sites in three replicates of colon cancer cell lines HT29 and SW480 with those detected in at least two replicates used for further analysis and highlighted (top), and their overlap between cell lines (bottom), represented using Venn diagram. (B) Heatmap representing p65 binding sites in the two cell lines. (C) Motifs highly enriched in p65 binding sites identified by HOMER using de novo motif analysis and sorted by p-value. (D) Genomic distribution of p65 binding sites in relation to gene locations. (E) Biological functions enriched in genes nearest p65 binding sites (-50kb +2kb).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 ZL0580 inhibits Tat transactivation and key factors in HIV transcription elongation. ( A and B ) Western blot measurement of Tat and NF-kappaB ( A ) and cellular proteins involved in transcription elongation ( B ) in WT J-Lat cells 24 hours after treatment. ( C ) Co-IP analysis for binding of CDK9 to Tat or BRD4 in WT J-Lat cells 24 hours after treatment. Control IgG Co-IP and input CDK9 were used as controls. Total/input CDK9 blots in panels ( B and C ) represent the same experiment. ( D ) ELL2 protein expression in WT and BRD4-KO J-Lat cells 24 hours after treatment. ( E ) ELL2 mRNA expression by qPCR in WT J-Lat cells 24 hours after treatment. ( F ) Effect of protease inhibition by MG132 on ELL2 protein levels in WT J-Lat cells. Cells were pretreated with proteasome inhibitor MG-132 for 6 hours (bottom) or not treated (top), followed by treatment with PMA or PMA+ZL0580 (10 muM) for 18 hours. ELL2 protein was measured by Western blot. ( G ) Phosphorylated RNAPII (Ser 2 CTD) in WT (top) and BRD4-KO (bottom) J-Lat cells after different treatments. Loading control GAPDH in panel ( D and G ) represents the same experiment. ( H and I ) ChIP-qPCR analysis for recruitment of Tat ( H ) or BRD4 ( I ) to HIV 5' LTR in PMA-activated or unstimulated J-Lat cells 24 hours after treatment. ChIP using control IgG was included for normalization. qPCR was conducted to quantify the precipitated HIV 5' LTR region. Data were normalized to NC. Error bars ( E ,

33-9900

Antibody data