SDS-polyacrylamide gel electrophoresis and WB were performed as described previously [14 (link)]. Antibodies used for analysis were anti-NXT1 rabbit polyclonal antibody (pAb) (Abnova, Taipei, Taiwan), anti-CRM1 mouse monoclonal antibody (mAb) (BD Biosciences, San Jose, CA, USA), anti-NUP153 rat mAb (Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-NUP98 rat mAb (Abcam, Cambridge, UK), anti-NUP62 mouse mAb (BD Biosciences), anti-E1B-AP5 (or hnRNP UL1) mouse mAb (Santa Cruz Biotechnology), anti-FLAG M2 mouse mAb (Sigma-Aldrich, Saint Louis, MO, USA), anti-FLAG rabbit mAb (Sigma-Aldrich), anti-HA mouse mAb (Medical and Biological Laboratories, Nagoya, Japan), anti-β-actin mouse mAb (Sigma-Aldrich), anti-WSN (influenza A/WSN/33) rabbit pAb (a gift from Dr. Kazufumi Shimizu, Nihon University School of Medicine, Tokyo, Japan), horseradish-peroxidase (HRP)-conjugated goat anti-mouse IgG (Amersham Biosciences, Uppsala, Sweden), or HRP-conjugated goat anti-rabbit IgG (Amersham Biosciences). The intensity of NP bands was quantitated by ImageJ 1.50 software (National Institutes of Health, Bethesda, MD, USA).
Anti ha mouse mab
Manufactured by Medical & Biological Laboratories
Sourced in United States, Japan
The Anti-HA mouse mAb is a monoclonal antibody that specifically binds to the hemagglutinin (HA) epitope tag. It is a useful tool for the detection and purification of HA-tagged proteins in various applications such as Western blotting, immunoprecipitation, and flow cytometry.
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Lab products found in correlation
2 protocols using anti ha mouse mab
1
SDS-PAGE and Western Blot Protocol
2
Quantitative Binding Assay of NP-CRM1
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NP-FLAG–immobilized agarose beads and purified CRM1-HA protein from transfected HEK293T cells were prepared as previously described (Chutiwitoonchai et al., 2014 (link)). Pull-down assays of NP/CRM1 interaction with DP2392-E10 were performed by incubation of 15 µl of NP-FLAG–immobilized agarose beads (50% slurry), 2 µg of purified CRM1-HA protein, and either DMSO or 3, 10, 30, or 100 µM DP2392-E10 in bind/wash buffer (10 mM Tris-Cl [pH 7.8], 150 mM NaCl, 0.05% NP-40) at 4 °C overnight. The beads were pulled down, washed five times with bind/wash buffer, and subjected to western blot analysis with anti-HA mouse mAb (Medical and Biological Laboratories, Nagoya, Japan) and anti-FLAG rabbit mAb (Sigma-Aldrich). The intensity of CRM1 and NP bands were quantitated by ImageJ 1.50 software (National Institute of Health, Bethesda, MD, USA).
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