NFKB1 anticorps (AA 740-964)

N° du produit ABIN6144571

Détail du produit anti-NFKB1 anticorps

Antigène: NFKB1 (Nuclear Factor of kappa Light Polypeptide Gene Enhancer in B-Cells 1 (NFKB1))

Épitope: AA 740-964

Reactivité: Humain

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp NFKB1 est non-conjugé

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunofluorescence (IF)

Séquence: AAGADPLVEN FEPLYDLDDS WENAGEDEGV VPGTTPLDMA TSWQVFDILN GKPYEPEFTS DDLLAQGDMK QLAEDVKLQL YKLLEIPDPD KNWATLAQKL GLGILNNAFR LSPAPSKTLM DNYEVSGGTV RELVEALRQM GYTEAIEVIQ AASSPVKTTS QAHSLPLSPA STRQQIDELR DSDSVCDSGV ETSFRKLSFT ESLTSGASLL TLNKMPHDYG QEGPL

Réactivité croisée: Humain, Souris, Rat

Attributs du produit: Polyclonal Antibodies

Immunogène: Recombinant fusion protein containing a sequence corresponding to amino acids 740-964 of human NFKB1 (NP_001158884.1).

Isotype: IgG

Information d'application

Indications d'application: WB,1:500 - 1:2000,IHC,1:50 - 1:200,IF,1:50 - 1:100

Commentaires:

HIGH QUALITY

Restrictions: For Research Use only

Independent Validation (IF)

Validation #104072 (Immunofluorescence)

by: Klinik für Anästhesiologie, Intensivmedizin und Schmerztherapie des Universitätsklinikums Knappschaftskrankenhaus Bochum

No.: #104072

Date: 21.11.2019

Antigène: NFKB1

Numéro du lot: 3560449004

Application validée: Immunofluorescence

Contrôle positif:

U-937 human monocytic cell line

Contrôle négative:

No primary antibody control

Conclusion:

Passed. ABIN6144571 specifically stains NF-κB1 in U-937 human monocytic cells.

Validation Images

U937 monocytic cells stained with with anti-NF-κB1/p50 antibody ABIN6144571 and a TRITC-labeled secondary antibody (bottom row, red). Counterstain with DAPI (blue). Negative control without primary antibody (top row) shows no specific TRITC signal.

Protocole

Anticorps primaire: ABIN6144571

Anticorps secondaire: TRITC-conjugated AffiniPure donkey anti-rabbit IgG antibody (611-025-215, Jackson ImmunoResearch Europe Ltd.)

Full Protocol:

• Grow U-937 cells in RPMI 1640 Medium (Gibco, 21875034) supplemented with 10% FCS (Gibco, A3840001) and Pen/Strep (Gibco, 15140122), at 37°C and 5% CO₂ in 300µL in 24-well plates (Sarstedt AG&Co. KG).
• Use cytospin to spin cells onto slide.
• Fix cells on coverslide in 4% paraformaldehyde (Carl Roth GmbH, 0335.1) for 30min at RT.
• Wash cells 3x for 5min with PBS (Sigma-Aldrich, 8537).
• Permeabilize cells in 100µL PBS containing 0.1 % Triton X-100 (Sigma-Aldrich, T8787) for 5min at RT.
• Wash cells 3x for 5min with PBS.
• Block non-specific binding with 1x Duolink Blocking solution (Sigma-Aldrich, DUO82007) for 30min at RT.
• Incubate cells with primary antibody rabbit anti-human NF-κ1/p50 (antibodies-Online, ABIN6144571, lot 3560449004) diluted 1:100 in 1x Duolink Antibody diluent (Sigma-Aldrich, DUO82008) ON at 4°C.
• Wash cells 3x for 5min with PBS.
• Incubate cells with secondary TRITC-conjugated AffiniPure donkey anti-rabbit IgG antibody (611-025-215, Jackson ImmunoResearch Europe Ltd.) diluted 1:400 in 1x Duolink Antibody diluent for 1h at RT in the dark.
• Wash cells 2x for 5min with PBS.
• Mount coverslips on glass slides in 50µL ProLong Gold antifade mountant with DAPI (Invitrogen, P36931).
• Image acquisition with an Olympus widefield microscope.

Notes:

Strong red staining dependent on the presence of primary antibody indicates that the antibody works. Negative control lacking the primary antibody shows only background fluorescence.

Stockage

Format: Liquid

Buffer: PBS with 0.02 % sodium azide,50 % glycerol, pH 7.3.

Agent conservateur: Sodium azide

Précaution d'utilisation: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Stock: -20 °C

Stockage commentaire: Store at -20°C. Avoid freeze / thaw cycles.

Références pour anticorps anti-NFKB1 (ABIN6144571)

Du, Qiao, Chen, Chen, Liu, Lin, Wang, Xie: "Toll-Like Receptor 4 Mediates Methamphetamine-Induced Neuroinflammation through Caspase-11 Signaling Pathway in Astrocytes." dans: Frontiers in molecular neuroscience, Vol. 10, pp. 409, (2017) (PubMed).

Ye, Chen, Li, Zhao, He, Zohaib, Song, Deng, Zhang, Chen, Cao: "Japanese Encephalitis Virus NS5 Inhibits Type I Interferon (IFN) Production by Blocking the Nuclear Translocation of IFN Regulatory Factor 3 and NF-κB." dans: Journal of virology, Vol. 91, Issue 8, (2017) (PubMed).

Détails sur NFKB1

Antigène: NFKB1 (Nuclear Factor of kappa Light Polypeptide Gene Enhancer in B-Cells 1 (NFKB1))

Autre désignation: NFKB1 (NFKB1 Produits)

Sujet: This gene encodes a 105 kD protein which can undergo cotranslational processing by the 26S proteasome to produce a 50 kD protein. The 105 kD protein is a Rel protein-specific transcription inhibitor and the 50 kD protein is a DNA binding subunit of the NF-kappa-B (NFKB) protein complex. NFKB is a transcription regulator that is activated by various intra- and extra-cellular stimuli such as cytokines, oxidant-free radicals, ultraviolet irradiation, and bacterial or viral products. Activated NFKB translocates into the nucleus and stimulates the expression of genes involved in a wide variety of biological functions. Inappropriate activation of NFKB has been associated with a number of inflammatory diseases while persistent inhibition of NFKB leads to inappropriate immune cell development or delayed cell growth. Alternative splicing results in multiple transcript variants encoding different isoforms, at least one of which is proteolytically processed.,NFKB1,CVID12,EBP-1,KBF1,NF-kB1,NF-kappa-B,NF-kappaB,NFKB-p105,NFKB-p50,NFkappaB,p105,p50,Epigenetics & Nuclear Signaling,Transcription Factors,Cancer,Signal Transduction,Cell Biology & Developmental Biology,Apoptosis,Inhibition of Apoptosis,Death Receptor Signaling Pathway,Immunology & Inflammation,B Cell Receptor Signaling Pathway,T Cell Receptor Signaling Pathway,IL-6 Receptor Signaling Pathway,NF-kB Signaling Pathway,Cell Intrinsic Innate Immunity Signaling Pathway,NFKB1

Poids moléculaire: 85 kDa/105 kDa

ID gène: 4790

UniProt: P19838

Pathways: Signalisation p53, Signalisation NF-kappaB, Signalisation RTK, TCR Signaling, Signalisation TLR, Fc-epsilon Receptor Signaling Pathway, Neurotrophin Signaling Pathway, Activation of Innate immune Response, Myometrial Relaxation and Contraction, Regulation of Carbohydrate Metabolic Process, Hepatitis C, Toll-Like Receptors Cascades, BCR Signaling, S100 Proteins