The following primary antibodies were used for western blotting: anti-IRP2 (Santa Cruz Biotechnology # 33682, 1:500; Cell Signaling # 37135, 1:500), anti-FBXL5 (NeoClone # N0036, 1:500; BioLegend # 672604, 1:500), anti-GAPDH (Cell Signaling # 2118, 1:1000; Santa Cruz Biotechnology # sc-47724, 1:5000), anti-CAND1 (Santa Cruz Biotechnology # 10672, 1:2000; Bethyl Laboratories # A302-901A, 1:1000), anti-CAND2 (Bethyl Laboratories # A304-046A, 1:1000), anti-CUL1 (Invitrogen # 32-2400, 1:500; Bethyl Laboratories # A303-373A, 1:500), anti-CUL2 (Thermo Fisher Scientific # 51-1800, 1:1000), anti-CUL3 (Cell Signaling # 2759, 1:500), anti-CUL4A (Cell Signaling # 2699, 1:1000), anti-BCL10 (Santa Cruz Biotechnology # sc-5273, 1:200), anti-SKP1 (Cell Signaling # 12248, 1:1000), anti-SKP2 (Cell Signaling # 2652, 1:1000), anti-HA (Cell Signaling # 3724, 1:1000), anti-FLAG (Sigma‒Aldrich # F1804, 1:5000), anti-StrepII (Abcam # ab76949, 1:500), and anti-IκBα (Abcam # ab32518, 1:1000). The secondary antibodies used for western blotting included Alexa Fluor 790-conjugated anti-mouse IgG (Thermo Fisher Scientific # ab186699, 1:10,000), Alexa Fluor 790-conjugated anti-rabbit IgG (Thermo Fisher Scientific # A11374, 1:10,000), Alexa Fluor 680-conjugated anti-rabbit IgG (Thermo Fisher Scientific # ab175772, 1:10,000), and Alexa Fluor 680-conjugated anti-mouse IgG (Abcam # A10038, 1:10,000).
Anti iκbα
Manufactured by Abcam
Sourced in United Kingdom, United States
About the product
Anti-IκBα is a lab equipment product that functions as an antibody. Its core purpose is to bind and detect the IκBα protein, which plays a role in the regulation of the NF-κB signaling pathway.
Automatically generated - may contain errors
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Get pricing insights and sourcing optionsSpelling variants (same manufacturer)
Anti-phospho-IκBα - 4 citations
Rabbit anti-phospho-IκBα - 3 citations
Anti-p-IκBα antibody - 3 citations
Anti-phosphorylated (p)-IκBα - 3 citations
Anti-IκBα (ab32518), - 2 citations
Rabbit polyclonal anti- IκBα - 2 citations
Rabbit anti-IκBα antibody - 2 citations
Rabbit monoclonal anti-IκBα - 2 citations
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The spelling variants listed below correspond to different ways the product may be referred to in scientific literature.
These variants have been automatically detected by our extraction engine, which groups similar formulations based on semantic similarity.
Product FAQ
66 protocols using «anti iκbα»
1
Western Blotting Antibody Protocol
2
Investigating NF-κB Signaling Pathways
Tumor cells were lysed in lysis buffer (20 mm Tris-HCl, pH 7.4, 150 mm NaCl, 1 mm EDTA, 1% Triton X-100) plus a protease inhibitor cocktail (Abcam, ab271306) for 60 min on ice. Proteins were separated on a 4–20% SDS-polyacrylamide gradient gel and transferred to an Immobilon-P transfer membrane (Millipore). The following antibodies were used after dilution with TBS containing 0.1% Tween 20 and 3% nonfat dry milk: anti-p65 (Abcam, ab16502), anti-phospho-p65 (Ser536) (CST, #3031), anti-IκBα (Abcam, ab97783), anti-IκBα phospho-Ser32 (CST, #9241), anti-Fas (Invitrogen, PA5–115214), anti-MCL-1 (CST, #4572), anti-BCL-2 (Abcam, ab59348), anti-BCL-6 (Abcam, ab19011), anti-BAX (Abcam, ab216494), anti-α-tubulin-acetyl-Lys40 (CST, #3971), anti-α-tubulin (Invitrogen, MA1–19162), anti-stathmin 1 (Abcam, ab131481), anti-phospho-stathmin 1 (S63) (Abcam, ab192648), anti-MAP4 (Bethyl Laboratories, #A301-489A), anti-phospho-MAP4 (S787) (GL Biochem), anti-IKKα + anti-IKKβ (GeneTex, GTX52348), anti-phospho-IKKα/β (Ser176/180) (CST, #2697), and anti-β-actin (Sigma-Aldrich, A5441) followed by incubation with an HRP-linked anti-rabbit IgG (CST, #7074) or anti-mouse IgG (Invitrogen, #31430). Immunodetection was performed using the Clarity ECL Western Blotting Substrates kit (BIO-RAD). For a co-immunoprecipitation experiment, MDS-L cells were treated with GNA, paclitaxel, and TN16 for 16 h at the indicated doses. After treatment, the cells were treated with 0.2% Triton X-100 for 5 min and then washed twice with phosphate-buffered saline. Whole cell lysates were prepared, and immunoprecipitation was performed with α-tubulin, IgG and GAPDH antibodies using equal amounts of proteins. After immunoprecipitation, Western blotting was performed using antibodies specific for α-tubulin, GAPDH, and IgG, and the co-immunoprecipitated NF-κB was detected using an anti-p65 antibody. Quantifications were done using ImageJ software.
3
Immunoblotting and ELISA for Macrophage Activation
Immunoblotting and detection of secreted bronectin were performed as previously described [69] [70] (link)[71] (link) . The primary antibodies were CD163 (Abcam, ab182422), CD68 (Abcam, ab125212), anti-α-tubulin (Sigma-Aldrich, T9026), anti-TGF-β1 (Abcam, ab215715), anti-bronectin (Sigma-Aldrich, F6140), anti-phospho-NF-κB (Cell Signaling Technology, 3033), anti-NF-κB (Cell Signaling Technology, 4764), anti-IκBα (Abcam, ab32518), anti-phospho-cJun (Cell Signaling Technology, 9164), anti-NLRP3 (Cell Signaling Technology, Danvers, MA, #15101), and anti-caspase1 (Abcam, ab207802). The intensity of each band was quanti ed using ImageJ software.
Enzyme-linked immunosorbent assay TGF-β1 protein in mice peritoneal uid samples, and TNF-α and IL-1β protein in THP-1 supernatant samples were measured with an enzyme-linked immunosorbent assay (ELISA) kit (R&D Systems, Minneapolis, MN) in accordance with the manufacturer's instructions. TNF-α and IL-1β concentrations were normalized to the total protein content of THP-1 cells. Figure 2
Enzyme-linked immunosorbent assay TGF-β1 protein in mice peritoneal uid samples, and TNF-α and IL-1β protein in THP-1 supernatant samples were measured with an enzyme-linked immunosorbent assay (ELISA) kit (R&D Systems, Minneapolis, MN) in accordance with the manufacturer's instructions. TNF-α and IL-1β concentrations were normalized to the total protein content of THP-1 cells. Figure 2
4
Western Blot Analysis of HCECs
Total proteins of HCECs were extracted using rapid lysis buffer (Solarbio, Beijing, China). Subsequently, lysates were collected and centrifuged at 4°C for 15 min, protein concentrations of supernatants were measured by BCA assay (Beyotime, Shanghai, China, cat. no. P0011), proteins were mixed with SDS loading buffer in a 4:1 ratio and then boiled for 10 min. Equal amounts of protein (20 μg) were loaded onto a 4%–20% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride (PVDF) membranes. Following blocking the membranes with 5% skimmed milk for 2 h at room temperature, and the membranes were incubated with primary antibodies at 4°C overnight. The following primary antibodies were used: Anti-Bax (1:1000; Abcam, Massachusetts, United States; cat. no. ab32503), anti-Bcl-2 (1:1000; Abcam, Massachusetts, United States cat. no. ab182858), anti-GAPDH (1:1000; Abcam, Massachusetts, United States; cat. no. ab181602), anti-TLR4 (1:1000; Abcam, Massachusetts, United States; cat. no. ab218987), anti-MyD88 (1:1000; Abcam, Massachusetts, United States; cat. no. ab133739), anti-Tirap (1:1000; Abcam, Massachusetts, United States; cat. no. ab17218), anti-IKK-α (1:1000; Abcam, Massachusetts, United States; cat. no. ab32041), anti-p-IKK-α (1:1000; Abcam, Massachusetts, United States; cat. no. ab38515), anti-NF-κB p65 (1:1000; Abcam, Massachusetts, United States; cat. no. ab207297), anti-p-NF-κB p65 (1:1000; Abcam, Massachusetts, United States; cat. no. ab239882), anti-IκB-α (1:1000; Abcam, Massachusetts, United States; cat. no. ab32518), anti-p-IκB-α (1:1000; Abcam, Massachusetts, United States; cat. no. ab92700). Then, they were incubated with HRP-conjugated Goat anti-rabbit IgG antibody (1:5000; Abcam, Massachusetts, United States; cat. no. ab6721) at room temperature for 2 h. Finally, each membrane was developed using a chemiluminescence (ECL) detection kit (Beyotime, Shanghai, China, cat. no. P0018S) and visualized using a chemiluminescence detection system (VILBER, Paris, France). The gray value of the target bands was calculated using Image. J software (version v1.8.0; National Institutes of Health).
5
Protein Extraction and Western Blot Analysis
The total proteins of the samples were extracted using lysis liquid (Lot No. P0013E, Beyotime Biotech. Inc., Shanghai, China) according to the procedures laid out by the manufacturer. Subsequently, the protein concentration was determined by applying a BCA kit (Lot No. KGP902 , KeyGEN BioTECH, Jiangsu, China). 10 % SDS - PAGE was used to separate the sample proteins, which were then transferred to a polyvinylidene difluoride membrane (Lot No. 1620177, Bio-Rad, Hercules, CA, USA), and TBST solution that contained 5 % milk was applied for blocking. After blocking for 1.5 h, the primary antibody was diluted with TBST solution to the following working concentrations: anti-MYDGF (myeloid cell–specific myeloid-derived growth factor, 1:1000, Proteintech, Lot No. 11353-1-AP, USA), anti-MAP4K4 (mitogen-activated protein kinase kinase 4, 1:1000, Cell Signaling Technology, Lot No. 3485 S, USA), anti-P-p65 (phosphorylation-nuclear factor NF-kappa-B p105 subunit, 1:1000, Abcam, Lot No. ab239882, England), anti-p65 (nuclear factor NF-kappa-B p105 subunit, 1:1000, Abcam, Lot No. ab32536, England), anti-P-IκBα (1:1000, Abcam, Lot No. ab133462, England), anti-IκBα (1:1000, Abcam, Lot No. ab32518, England), anti-PKC (protein kinase C, 1:1000, Abcam, Lot No. ab181558, England), anti-IL6 (1:1000, Wuhan Servicebio Technology Co., Ltd., Lot No. GB11117, China), anti-TNF-α (1:1000, Wuhan Sanying Co., Ltd., Lot No.17590-1-ap, China), and anti-β-actin (1:1000, Cell Signaling Technology, Lot No. 4967 S, USA) at 4 °C overnight. Next, the membrane was rinsed with TBST three times and incubated with a secondary antibody (1:2000, Lot No. S0001 or Lot No. S0002, Affinity Biosciences company, Jiangsu, China) for 1 h at room temperature. Finally, they were exposed using an ECL reagent kit (Thermo Science, New Jersey, USA), and the bands were visualized and analyzed using Image-Pro Plus 6.0 software (Media Cybernetics, Bethesda, USA) and normalized to the band intensities of β-actin. All experiments were repeated three times.
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