Human/Mouse IkB-alpha Antibody

Detection of Human/Mouse IkB-alpha by Western Blot. Western blot shows lysates of NIH-3T3 mouse embryonic fibroblast cell line, DA3 mouse myeloma cell line, LNCaP human prostate cancer cell line, and Daudi human Burkitt's lymphoma cell line. PVDF membrane was probed with 0.1 µg/mL of Sheep Anti-Human/Mouse IkB-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4299) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for IkB-a at approximately 42 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

IkB‑ alpha in HeLa Human Cell Line. IkB-a was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human/Mouse IkB-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4299) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

IkB‑ alpha in HeLa Human Cell Line. IkB-a was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human/Mouse IkB-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4299) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; NL010) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Western Blot Shows Human IkB‑ alpha Specificity by Using Knockout Cell Line. Western blot shows lysates of HEK293T human embryonic kidney parental cell line and IkB-a knockout HEK293T cell line (KO). PVDF membrane was probed with 0.1 µg/mL of Sheep Anti-Human/Mouse IkB-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4299) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for IkB-a at approximately 38 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of IkB‑ alpha in RAW 264.7 Mouse Monocyte/Macrophage Cell Line. IkB‑ alpha was detected in immersion fixed RAW 264.7 Mouse Monocyte/Macrophage Cell Line using Sheep Anti-Human/Mouse IkB‑ alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4299) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.