nemo antiserum (Cell Signaling Technology Inc)

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Cell Signaling Technology Inc nemo antiserum

The 9SG mutation abolishes the ability of <t>NEMO</t> to function in signal-induced activation of IκB kinase. (A) The indicated FLAG-tagged NEMO constructs were transfected into 293T cells, and extracts were immunoprecipitated (IP) with anti-FLAG beads. Immunoprecipitates were then analyzed by anti-FLAG (bottom) and anti-IKKβ (top) Western blotting. In the Input lanes, 4% of the extract used in the immunoprecipitations was analyzed by Western blotting. (B) Whole cell extracts from wild-type and NEMO-knockout 293T cells were analyzed by anti-NEMO Western blotting (top) or Ponceau staining for total protein (bottom). (C) 293T NEMO knockout cells were transfected with plasmids for the expression of 7XAla- or 9SG-NEMO. Cells that were either untreated (−) or treated with TNFα (+) were analyzed by Western blotting <t>for</t> <t>phospho-IκBα</t> or NEMO or by Ponceau staining. (D) Mouse NEMO knockout fibroblasts were stably transduced with retroviral vectors for the indicated NEMO proteins. Stable cell lines were then untreated (−) or treated (+) with the indicated compounds. Extracts were analyzed by Western blotting for the indicated proteins.

Nemo Antiserum, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 94 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/nemo+antiserum/IKKgamma+Antibody/pmc09295417-169-30-34
Average 93 stars, based on 94 article reviews

nemo antiserum - by Bioz Stars, 2026-10

93/100 stars

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1) Product Images from "A Central Region of NEMO is Required for IKKβ-Induced Conformational Change and for Signal Propagation"

Article Title: A Central Region of NEMO is Required for IKKβ-Induced Conformational Change and for Signal Propagation

Journal: Biochemistry

doi: 10.1021/acs.biochem.8b01316

The 9SG mutation abolishes the ability of NEMO to function in signal-induced activation of IκB kinase. (A) The indicated FLAG-tagged NEMO constructs were transfected into 293T cells, and extracts were immunoprecipitated (IP) with anti-FLAG beads. Immunoprecipitates were then analyzed by anti-FLAG (bottom) and anti-IKKβ (top) Western blotting. In the Input lanes, 4% of the extract used in the immunoprecipitations was analyzed by Western blotting. (B) Whole cell extracts from wild-type and NEMO-knockout 293T cells were analyzed by anti-NEMO Western blotting (top) or Ponceau staining for total protein (bottom). (C) 293T NEMO knockout cells were transfected with plasmids for the expression of 7XAla- or 9SG-NEMO. Cells that were either untreated (−) or treated with TNFα (+) were analyzed by Western blotting for phospho-IκBα or NEMO or by Ponceau staining. (D) Mouse NEMO knockout fibroblasts were stably transduced with retroviral vectors for the indicated NEMO proteins. Stable cell lines were then untreated (−) or treated (+) with the indicated compounds. Extracts were analyzed by Western blotting for the indicated proteins.

Figure Legend Snippet: The 9SG mutation abolishes the ability of NEMO to function in signal-induced activation of IκB kinase. (A) The indicated FLAG-tagged NEMO constructs were transfected into 293T cells, and extracts were immunoprecipitated (IP) with anti-FLAG beads. Immunoprecipitates were then analyzed by anti-FLAG (bottom) and anti-IKKβ (top) Western blotting. In the Input lanes, 4% of the extract used in the immunoprecipitations was analyzed by Western blotting. (B) Whole cell extracts from wild-type and NEMO-knockout 293T cells were analyzed by anti-NEMO Western blotting (top) or Ponceau staining for total protein (bottom). (C) 293T NEMO knockout cells were transfected with plasmids for the expression of 7XAla- or 9SG-NEMO. Cells that were either untreated (−) or treated with TNFα (+) were analyzed by Western blotting for phospho-IκBα or NEMO or by Ponceau staining. (D) Mouse NEMO knockout fibroblasts were stably transduced with retroviral vectors for the indicated NEMO proteins. Stable cell lines were then untreated (−) or treated (+) with the indicated compounds. Extracts were analyzed by Western blotting for the indicated proteins.

Techniques Used: Mutagenesis, Activation Assay, Construct, Transfection, Immunoprecipitation, Western Blot, Knock-Out, Staining, Expressing, Stable Transfection, Transduction

Western Blot:

Article Title: A Central Region of NF-κB Essential Modulator Is Required for IKKβ-Induced Conformational Change and for Signal Propagation.
Article Snippet: Subscriber access provided by UNIV OF SOUTHERN INDIANA is published by the American Chemical Society.. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society.. Copyright © American Chemical Society.

Incubation:

Article Title: A Central Region of NEMO is Required for IKKβ-Induced Conformational Change and for Signal Propagation
Article Snippet: .. In all cases, samples containing approximately equal amounts of protein were separated on SDS-polyacrylamide gels, proteins were transferred to nitrocellulose membranes, and filters were incubated overnight at 4 °C with NEMO antiserum (cat. #2685, Cell Signaling Technology; 1:1000 dilution) or with phospho-IκBα antiserum (cat. #9246, Cell Signaling Technology; 1:1000 dilution). .. Horseradish peroxidase-conjugated secondary antiserum was added, and immunoreactive proteins were detected by SuperSignal Dura West chemiluminescence (Thermo Fisher Scientific).

other:

Article Title: A Central Region of NEMO is Required for IKKβ-Induced Conformational Change and for Signal Propagation
Article Snippet: For analyzing phosphorylation of IκBα, cells were first treated with 20 ng/mL TNFα (R&D Systems) for 10 min, 2 ng/mL of IL-1 for 6 min, or 100 ng/mL of LPS for 12 min.

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