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FIGURE: 1 / 5
IkB alpha Antibody (740061TP647) in WB
Western blot was performed using IkB alpha Recombinant Rat Monoclonal Antibody (MFRDTRK), Alexa FluorTM Plus 647 (Product # 740061TP647, 1:400) and a ~36 kDa band corresponding to IkB alpha was observed across cell lines tested. Whole cell extracts (30 µg lysate) of NIH/3T3 (Lane 1), THP-1 (Lane 2), LNCaP (Lane 3), Hep G2 (Lane 4), and HeLa (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot™ 2 Dry B... View More
Please note: We are reviewing Western blot images included in the antibody testing data in our catalog, including those provided by third parties. Unless expressly labeled or annotated as “raw-unedited”, Western blot images included in the antibody testing data in our catalog may have been edited, optimized or otherwise adjusted for presentation.
Product Details
740061TP647
Applications
Tested Dilution
Publications
Product Specifications
Species Reactivity
Human
Host/Isotype
Rat
/ IgG2b, kappa
Expression System
Expi293
Class
Recombinant Monoclonal
Type
Antibody
Clone
MFRDTRK
Conjugate
Alexa Fluor™ Plus 647
Alexa Fluor™ Plus 647
Alexa Fluor™ Plus 647
Excitation/Emission Max
658/675 nm
View spectra
Form
Liquid
Concentration
1.0 mg/mL
Amount
50 µg
Purification
Affinity chromatography
Storage buffer
proprietary buffer, pH 6.8
Contains
0.008% Bromonitrodioxane, 0.008% Methylisothiazolone
Storage conditions
4°C, store in dark, DO NOT FREEZE!
Shipping conditions
Wet ice
RRID
Product Specific Information
Alexa Fluor™ Plus recombinant antibodies are conjugated using new, proprietary dye chemistry so you can generate stunning data. Alexa Fluor™ Plus antibodies represent an advancement in fluorescent conjugate technology. Alexa Fluor™ Plus antibodies provide brighter signal compared to leading Alexa Fluor™ antibodies, providing you with better signal-to-noise for your critical experiments. These antibodies show better specificity and lot-to-lot consistency as these are recombinant antibodies, generated by cloning specific genes for the desired antibodies into an expression vector and expressed in vitro.
Using conjugate solutions: Centrifuge the protein conjugate solution briefly in a microcentrifuge before use; add only the supernatant to the experiment. This step will help eliminate any protein aggregates that may have formed during storage, thereby reducing nonspecific background staining.
Applications Tested: This MFRDTRK antibody has been tested by western blotting and flow cytometric analysis of HeLa cells treated with TNF alpha. This may be used for western blotting at 5 µg/mL and for flow cytometry at less than or equal to 2 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Excitation: 658 nm; Emission: 675 nm; Laser: Red Laser
Filtration: 0.2 µm post-manufacturing filtered.
Target Information
IkB-alpha is a 40 kDa protein that functions to inhibit NF- kappaB activity. The inhibition occurs via protein-protein interaction between I kappaB proteins and NF- kappaB dimers in the cytosol. The interaction of I kappa B-alpha with NF- kappaB masks the nuclear localization sequence of NF- kappaB, preventing NF- kappaB translocation to the nucleus. A variety of stimuli can activate gene expression by liberating NF- kappaB through the degradation of I kappaB alpha. These stimuli include the proinflammatory cytokines TNF- alpha and IL-1 beta, chemokines, PMA, growth factors, LPS, UV irradiation, viral infection, as well as various chemical and physical stresses. In humans, the gene is located on the q arm of chromosome 14. Activation of NFkB requires that IkB be phosphorylated on specific serine residues, which results in targeted degradation of IkB. IkB kinase alpha (IKK alpha), previously designated CHUK, interacts with IkB-alpha and specifically phosphorylates IkB-alpha on the sites that trigger its degradation Serines 32 and 36. IKK alpha appears to be critical for NFkB activation in response to proinflammatory cytokines. Phosphorylation of IkB by IKK alpha is stimulated by the NFkB inducing kinase (NIK), which itself is a central regulator for NFkB activation in response to TNF and IL-1. The functional IKK complex contains three subunits, IKK alpha, IKK beta and IKK gamma, and each appear to make essential contributions to IkB phosphorylation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Bioinformatics
Protein Aliases: I-kappa-B-alpha; IkappaBalpha; ikB-alpha; IkBa; IKBalpha; MAD3; major histocompatibility complex enhancer-binding protein MAD3; Mutations: 945:OK; nuclear factor of kappa light chain gene enhancer in B-cells; nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha; unnamed protein product
Gene Aliases: EDAID2; IKBA; MAD-3; NFKBI
UniProt ID: (Human) P25963
Entrez Gene ID: (Human) 4792
negative regulation of transcription from RNA polymerase II promoter
I-kappaB kinase/NF-kappaB signaling
negative regulation of macrophage derived foam cell differentiation
negative regulation of lipid storage
negative regulation of cholesterol transport
tumor necrosis factor-mediated signaling pathway
toll-like receptor 4 signaling pathway
NIK/NF-kappaB signaling
negative regulation of protein import into nucleus
negative regulation of I-kappaB kinase/NF-kappaB signaling
positive regulation of transcription from RNA polymerase II promoter
positive regulation of inflammatory response
B cell receptor signaling pathway
positive regulation of transcription initiation from RNA polymerase II promoter
cellular response to cold
interleukin-1-mediated signaling pathway
cellular response to cytokine stimulus
negative regulation of cytokine production involved in inflammatory response
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