Ab138449

Manufactured by Abcam

Ab138449 is a primary antibody for the detection of a specific target protein. It is intended for use in various immunoassay applications, such as Western blotting, immunohistochemistry, and ELISA.

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Lab products found in correlation

Ripa buffer, by Thermo Fisher Scientific (4 mentions) Ab68481, by Abcam (2 mentions) Ab205718, by Abcam (2 mentions) Ab9385, by Abcam (2 mentions) Halt protease and phosphatase inhibitor cocktail, by Thermo Fisher Scientific (2 mentions) Goat serum, by Thermo Fisher Scientific (1 mentions) Paraformaldehyde (pfa), by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Triton x 100, by Merck Group (1 mentions) Ab25950, by Abcam (1 mentions) Ab14686, by Abcam (1 mentions) Ab139729, by Abcam (1 mentions) Ab203043, by Abcam (1 mentions) Ab229987, by Abcam (1 mentions) Ab178945, by Abcam (1 mentions) Ab62352, by Abcam (1 mentions) Non protein blocking solution, by Sangon (1 mentions)

4 protocols using ab138449

Western Blot Analysis of IRF3

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The antibodies used in this study were anti-IRF3 (Abcam, ab68481), anti-IRF3 (Phospho S396) (Abcam, ab138449), anti-GAPDH (Abcam, ab9385), and goat anti-rabbit IgG H&L (HRP) (Abcam, ab205718). Cells were harvested and lysed in radio-immunoprecipitation assay (RIPA) buffer (Thermo Scientific, catalog no. 89900) supplemented with Halt protease and phosphatase inhibitor cocktail (Thermo Scientific, catalog no. 78440) on ice. The cell lysates were subject to western blotting. GAPDH was used as a loading control.

Si L., Bai H., Oh C.Y., Jiang A., Hong F., Zhang T., Ye Y., Jordan T.X., Logue J., McGrath M., Belgur C., Calderon K., Nurani A., Cao W., Carlson K.E., Prantil-Baun R., Gygi S.P., Yang D., Jonsson C.B., tenOever B.R., Frieman M, & Ingber D.E. (2022). Self-assembling short immunostimulatory duplex RNAs with broad-spectrum antiviral activity. Molecular Therapy. Nucleic Acids, 29, 923-940.

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Western Blot Analysis of IRF3 Activation

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The antibodies used in this study were anti-IRF3 (Abcam, ab68481), anti-IRF3 (Phospho S396) (Abcam, ab138449), anti-GAPDH (Abcam, ab9385), and Goat anti-Rabbit IgG H&L (HRP) (Abcam, ab205718). Cells were harvested and lysed in RIPA buffer (Thermo Scientific, Cat#89900) supplemented with Halt^™ protease and phosphatase inhibitor cocktail (Thermo Scientific, Cat#78440) on ice. The cell lysates were subject to western blotting. GAPDH was used as a loading control.

Si L., Bai H., Oh C.Y., Zhang T., Hong F., Jiang A., Ye Y., Jordan T.X., Logue J., McGrath M., Belgur C., Nurani A., Cao W., Prantil-Baun R., Gygi S.P., Powers R.K., Frieman M., tenOever B.R, & Ingber D.E. (2021). Self-assembling short immunostimulatory duplex RNAs with broad spectrum antiviral activity. bioRxiv.

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Immunofluorescent Localization of Phospho-IRF3

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Cells were rinsed with PBS, fixed with 4% paraformaldehyde (Alfa Aesar) for 30 min, permeabilized with 0.1% Triton X-100 (Sigma-Aldrich) in PBS (PBST) for 10 min, blocked with 10% goat serum (Life Technologies) in PBST for 1 h at room temperature, and incubated with anti-IRF3 (Phospho S396) (Abcam, ab138449) antibody diluted in blocking buffer (1% goat serum in PBST) overnight at 4 °C, followed by incubation with Alexa Fluor 488 conjugated secondary antibody (Life Technologies) for 1 h at room temperature; nuclei were stained with DAPI (Invitrogen) after secondary antibody staining. Fluorescence imaging was carried out using a confocal laser-scanning microscope (SP5 X MP DMI-6000, Germany) and image processing was done using Imaris software (Bitplane, Switzerland).

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Western Blot Analysis of Protein Targets

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Tissue lysates were prepared using Cell Lysis Buffer for Western and IP (Biotime Biotechnology, Xiamen, Fujian, PRC). Protein concentrations were detected using Enhanced BCA Protein Assay Kit (Biotime Biotechnology). Equal amounts of proteins from each sample were subjected to SDS-PAGE followed by transfer of proteins to nitrocellulose membranes. Membranes were blocked with a non-protein blocking solution (Sangon Biotech) and incubated with a primary antibody overnight at 4 °C. After washing with TBST, membranes were incubated with a secondary antibody linked to Horseradish peroxidase (HRP). The blots were then developed with an ECL detection system as per the manufacturer's instructions. Rabbit anti-SGLT1 (ab14686), anti-PEPT1 (ab203043), anti-P47 phox (ab74095), anti-Nrf2 (ab62352), anti-Keap1 (ab139729), anti-iNOS (ab178945), anti-MR (ab229987), anti-p-IRF3 (Ser396)(ab138449), anti-IRF3 (ab25950), monoclonal antibodies were obtained from Abcam. Rabbit anti-Arg (93668S), anti-pSTAT1(Tyr701)(9167S), anti-STAT1 (9172T), anti-STAT6 (5397S), anti-STAT3 (30835S) monoclonal antibodies were obtained from CST (Shanghai, PRC). Mouse anti-βactin monoclonal antibody was obtained from Biotime Biotechnology.

Cao X., Tang L., Zeng Z., Wang B., Zhou Y., Wang Q., Zou P., & Li W. (2020). Effects of probiotic BaSC06 on intestinal digestion and absorption function, antioxidant capacity and macrophage polarization and microbiota composition in fattening pigs.

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