Human SCD-1 Antibody

Detection of Human SCD‑1 by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line and SK-Mel-28 human malignant melanoma cell line. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human SCD-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7550) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for SCD-1 at approximately 38 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Western Blot Shows Human SCD‑1 Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and SCD-1 knockout HeLa cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human SCD-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7550) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for SCD-1 at approximately 41 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Western Blot Shows Human SCD-1 Specificity Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and SCD-1 knockout HeLa cell line (KO). Nitrocellulose membrane was probed with 1 µg/mL of Sheep Anti-Human SCD-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7550) followed by HRP-conjugated anti-sheep IgG Secondary Antibody. A specific band was detected for SCD-1 at approximately 36 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.

SCD-1 Specificity is Shown by Immunocytochemistry in Knockout Cell Line. HeLa WT and SCD-1 KO cells were labelled with a green or a far-red fluorescent dye, respectively. Cells were stained with Sheep Anti-Human SCD-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7550) followed by incubation with an anti-sheep Alexa-fluor 555 coupled secondary antibody (upper panel). DAPI-only counterstained cells shown on a lower panel. Acquisition of the blue (nucleus-DAPI), green (identification of WT cells), red (antibody staining) and far-red (identification of KO cells) channels was performed. Representative images of the blue and red (grayscale) channels are shown. WT and KO cells are outlined with green and magenta dashed line, respectively. Primary antibody concentration used: 1 µg/mL. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).

Detection of SCD-1 by Immunoprecipitation. PMA-treated HeLa lysates were prepared and immunoprecipitation was performed using 2.0 µg of Sheep Anti-Human SCD-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7550) pre-coupled to Dynabeads protein G. Immunoprecipitated SCD-1 was detected with Mouse Anti-SCD-1 Antibody.The Ponceau stained transfer of the blot is shown. SM=4% starting material; UB=4% unbound fraction; IP=immunoprecipitate; HC=antibody heavy chain. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).