DMC (Figure 1A(Fig. 1) ), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and phosphate-buffered saline (PBS) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were obtained from Invitrogen-Gibco (Grand Island, NY, USA). Enzyme-linked immunosorbent assay (ELISA) kits for PGE2, TNF-α, IL-1ß, and IL-6 were purchased from R&D Systems, Inc. (St. Louis, MO, USA). Anti-iNOS (mouse), Anti-COX-2 (rabbit), Anti-IκB-α (mouse), anti-phosphorylated IκB-α (rabbit, anti-p-IκB-α), anti-c-Jun N-terminal kinase (rabbit, JNK), anti-phosphorylated JNK (rabbit, anti-p-JNK), anti-extracellular signal-regulated kinases (rabbit, ERK1/2), anti-phosphorylated ERK1/2 (rabbit, anti-p-ERK1/2), anti-p38 (rabbit), and anti-phosphorylated p38 (rabbit, anti-p-p38) mouse or rabbit antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). All other reagents were purchased from the Sigma-Aldrich Chemical Co. (St. Louis, MO, USA).
Anti phosphorylated iκbα
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-phosphorylated IκBα is a laboratory reagent used to detect the phosphorylated form of the IκBα protein. IκBα is a key regulator of the NF-κB signaling pathway and its phosphorylation is a critical step in the activation of NF-κB.
Automatically generated - may contain errors
Lab products found in correlation
Anti iκbα, by Cell Signaling Technology (7 mentions)
Anti phosphorylated erk1 2, by Cell Signaling Technology (3 mentions)
Anti phosphorylated jnk, by Cell Signaling Technology (3 mentions)
Anti phosphorylated p38, by Cell Signaling Technology (3 mentions)
Anti p38, by Cell Signaling Technology (3 mentions)
Anti erk1 2, by Cell Signaling Technology (2 mentions)
Anti jnk, by Cell Signaling Technology (2 mentions)
Anti p65, by Cell Signaling Technology (2 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Erk1 2, by Cell Signaling Technology (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Merck Group (1 mentions)
Enzyme linked immunosorbent assay elisa kits for pge2, by R&D Systems (1 mentions)
Interleukin 6 (il 6), by R&D Systems (1 mentions)
Tnf α, by R&D Systems (1 mentions)
Anti c jun n terminal kinase, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase conjugated goat anti mouse igg antibody, by Merck Group (1 mentions)
Anti akt, by Cell Signaling Technology (1 mentions)
Anti phosphoinositide 3 kinases pi3k, by Cell Signaling Technology (1 mentions)
Anti vegfr 2, by Merck Group (1 mentions)
10 protocols using anti phosphorylated iκbα
1
Anti-inflammatory Mechanisms of Bioactive Compounds
2
Measuring Signaling Pathways in Lung Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The protein extracts of lung were incubated with the primary antibody [anti-phosphoinositide 3-kinases (PI3K) (1:1000; Cell Signaling Technology); anti-nuclear factor kappa B (NFκB, p65) (1:200; Santa Cruz Biotechnology, Santa Cruz, CA, U.S.A.); anti-nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha (IκBα) (1:10000; Abcam plc); anti-phosphorylated IκBα (1:1000; Cell Signaling Technology); anti-VEGF (1:1000; Santa Cruz Biotechnology); anti-VEGFR-1, -phosphorylated VEGFR-1 (1:1000; Abcam plc); anti-VEGFR-2 (1:500; Millipore Corporation); anti-phosphorylated VEGFR-2 (1:1000; Cell Signaling Technology); anti-Rho-associated kinase (RhoA) (1:1000; Cell Signaling Technology); anti-Akt (1:500, Cell Signaling Technology); anti-phosphorylated Akt (1:2000, Cell Signaling Technology); anti-extracellular signal-regulated kinase (ERK), -phosphorylated ERK (1:3000, Millipore Corporation)]. Then the blots were incubated with the secondary antibody (horseradish peroxidase-conjugated goat anti-mouse IgG antibody, Sigma Chemical Co., St. Louis, MO, U.S.A.). With a computer assisted video densitometer and digitalized software (Kodak Digital ScienceTM ID Image Analysis Software, Eastman Kodak Co., Rochester, NY, U.S.A.), the blots were scanned, photographed then the signal intensity (integral volume) of the appropriate bands were analyzed.
3
Immunoblotting Analysis of Apoptosis Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Anti-PARP, anti-β-actin, anti-caspase-3, anti-caspase-9, VDAC, anti-phosphorylated ERK1/2, anti-phosphorylated p38, anti-phosphorylated JNK, anti-phosphorylated IκBα, anti-ERK1/2, anti-p38, anti-JNK, anti-Bcl-2, and anti-IκBα were purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA). α-Tubulin, anti-Bax (6A7) and His-probe were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against cytochrome c (for immunofluorescence, clone 6H2.B4; for western blot analysis, clone 7H8.2C12) were acquired from BD Pharmingen (San Diego, CA, USA), and the anti-cytochrome oxidase subunit IV (COX IV) antibody was purchased from Abcam (Cambridge, UK). Dichlorodihydrofluorescein diacetate (H2DCFDA), MitoSOX, DAPI, and DiOC6 were obtained from Molecular Probes (Eugene, OR, USA) and z-VAD-fmk, N-acetyl-cysteine (NAC), horseradish peroxidase-conjugated anti-mouse IgG and anti-rabbit IgG, and specific inhibitors of ERK (U0126), p38 mitogen-activated protein kinase (MAPK; SB203580), JNK (SP600125), and nuclear factor-κB (NF-κB; BAY 11-7082) were purchased from Calbiochem (San Diego, CA, USA). Anti-F4/80 and anti-CD11b were performed as recommended by the manufacturer (eBioscience, San Diego, CA, USA).
4
Comprehensive Western Blotting Procedure
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The western blotting procedure was followed from a previous study45 (link). The antibodies used in the present study included anti-DENV NS2B (1:3000; GeneTex, Irvine, CA, USA), anti-GAPDH (1:3000; GeneTex, Irvine, CA, USA), anti-COX-2 (1:1000, Cayman, ML, USA), and anti-Myc (1:2000; Abcam, Cambridge, MA, USA). In addition, anti-phosphorylated IκBα, IKKα/β, NF-κB, p38, JNK, and ERK and anti-total IκBα, IKKα, IKKβ, NF-κB, p38, JNK, and ERK antibodies were used (1:1000; Cell Signaling Technology, Inc. Danvers, MA, USA).
5
Western Blot Analysis of Dpep2 and NF-κB
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Heart homogenates or cells lysates were subjected to 10% sodium dodecyl polyacrylamide gel electrophoresis separation. The membranes were probed with the primary antibodies anti-Dpep2 (abcam), anti-phosphorylated-p65 (p-p65), anti-p65, anti-IκB-α, anti-phosphorylated-IκB-α, anti-Ikk-α, anti-phosphorylated-Ikk-α (p-Ikk-α) (Cell Signaling Technology) and anti-GAPDH (Sigma). HRP-conjugated anti-rabbit IgG antibody (Southern Biotech) was used as the secondary antibody. Detection was performed by enhanced cheminescence (Pierce; Thermo Fisher Scientific) and band intensities were quantified by ImageJ software.
6
Modulating T-cell Activation and Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies against human CD3 and CD28 for the stimulation of T cells were purchased from BioXcell (West Lebanon, NH, USA). Human TNFα recombinant protein was obtained from PeproTech EC Ltd. (London, UK). TRIZOL reagent for RNA isolation, MTT powder (1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan), and radioimmunoprecipitation assay (RIPA) buffer for cell lysis were provided by Sigma Chemical Co. (St. Louis, MO, USA). Staining reagents for AnnexinV (green) and caspase3/7 (red) were purchased from Essen Bio (Ann Arbor, MI, USA). SYBR Premix Ex Taq was obtained from TaKaRa (Shiga, Japan). Anti-p65, anti-LaminB, anti-IκBα, anti-phosphorylated IκBα, anti-phosphorylated ERK, anti-phosphorylated p38, anti-p38, anti-phosphorylated JNK, and anti-JNK were provided by Cell Signaling Technology (Danvers, MA, USA). Antibodies against actin and ERK were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). ECL Western blotting detection reagents were obtained from Thermo Fisher Scientific (Waltham, MA, USA). The RT PreMix kit was provided by Enzynomics (Daejeon, Korea). DSS was purchased from MP Biomedicals (Irvine, CA, USA).
7
Immunoblot Analysis of NMO Signaling Pathway
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were serum-starved overnight prior to stimulation with NMO IgG or control IgG then lysed in RIPA buffer containing protease/phosphatase inhibitors. Cell lysates (10–30 μg) were run on 4–15 % Criterion Tris–HCl gels (Biorad). After transfer, blots were probed using anti-IκB-α (Cell Signaling 9242), anti-phosphorylated IκB-α (Cell Signaling 2895), anti-p65 (Cell Signaling 8242), anti-NUP98 (Cell Signaling 2598), or anti-tubulin (Sigma T9026) antibodies.
8
Antibody Characterization for Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used: anti-His (ab9136 or ab18184, Abcam or sc-804, Santa Cruz), anti-Tax1 (Tab172, NIH), anti-p62 (GP62-C, Progen), anti-OPTN (100000, Cayman), anti-GM130 (610823, BD), anti-IKKγ (611306, BD), anti-phosphorylated IKKα/β (2078, Cell Signaling Technology), anti-IκBα (4814, Cell Signaling Technology), anti-phosphorylated IκBα (9246, Cell Signaling Technology), anti-FLAG (M2, Sigma-Aldrich), anti-Myc (4A6, Millipore), anti-Ub (P4D1, sc-8017, Santa Cruz), anti-actin (AC-74, Sigma-Aldrich), anti-Nup153 (147050002, Covance). HRP-Streptavidin was from Sigma (RAB-HRP3). For control immunoprecipitation, normal guinea pig IgG (sc-2711, Santa-Cruz) were used. For inhibition of lysosomal degradation, E64D and pepstatin (Sigma) were both used at 10 µg/mL.
9
Inflammatory Signaling Pathway Reagents
10
Western Blot Antibody Panel
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following primary antibodies were used: anti-p38, anti-phosphorylated p38, anti-ERK1/2, anti-phosphorylated ERK1/2, anti-IκBα, anti-phosphorylated IκBα, anti-P65, anti-phosphorylated P65, anti-Tak1, and anti-myc antibody were from Cell Signaling Technology. Anti-phosphorylated Rip2 was from Abgent. Anti-JNK antibody, anti-phosphorylated JNK antibody, and anti-phosphorylated Tak1 were from Abcam. Anti-Rip2 antibody was from Abnova (H0000876-M02). Anti-LRRK2 antibody was from Epitomics (MJFF2 c41-2). Anti-actin antibody was from Sigma (A5441). Anti-GFP antibody was from Thermo Fisher Scientific (A-11120).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!