Human Rab27a Antibody

Detection of Human Rab27a by Western Blot. Western blot shows lysates of human prostate tissue. PVDF membrane was probed with 2 µg/mL of Rabbit Anti-Human Rab27a Monoclonal Antibody (Catalog # MAB7245) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for Rab27a at approximately 26 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human Rab27a by Simple Western^TM. Simple Western shows lysates of Exosome Standards (K562) (NBP2-49864) and SK‑Mel‑28 human malignant melanoma cell line, loaded at 0.5 mg/ml. A specific band was detected for Rab27a at approximately 32 kDa (as indicated) using 20 µg/mL of Rabbit Anti-Human Rab27a Monoclonal Antibody (Catalog # MAB7245). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Rab27a in SK‑Mel‑28 Human Cell Line. Rab27a was detected in immersion fixed SK-Mel-28 human malignant melanoma cell line (left panel; positive staining) and Daudi human Burkitt's lymphoma cell line (right panel; negative staining) using Rabbit Anti-Human Rab27a Monoclonal Antibody (Catalog # MAB7245) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human Rab27a by Simple Western^TM. Simple Western lane view shows lysates of K562 human chronic myelogenous leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for Rab27a at approximately 31 kDa (as indicated) using 20 µg/mL of Rabbit Anti-Human Rab27a Monoclonal Antibody (Catalog # MAB7245). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Rab27a Specificity is Shown by Immunocytochemistry in Knockdown Cell Line. U‑87 MG human glioblastoma/astrocytoma parental cell line ctrl and Rab27a U-87 MG KD cells were labelled with a green or a far-red fluorescent dye, respectively. Cells were stained with Rabbit Anti-Human Rab27a Monoclonal Antibody (Catalog # MAB7245) followed by incubation with an Alexa-fluor 555 conjugated secondary antibody (upper panel). DAPI-only counterstained cells shown on a lower panel. Acquisition of the blue (nucleus-DAPI), green (identification of ctrl cells), red (antibody staining) and far-red (identification of KD cells) channels was performed. Representative images of the blue and red (grayscale) channels are shown. Ctrl and KD cells are outlined with green and magenta dashed line, respectively. Primary antibody concentration used: 3.33 µg/mL. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).

Western Blot Shows Rab27a Specificity Using Knockdown Cell Line. Western blot shows lysates of U‑87 MG human glioblastoma/astrocytoma parental cell line and Rab27a knockdown U-87 MG cell line (KD). Nitrocellulose membrane was probed with 2.5 µg/mL Rabbit Anti-Human Rab27a Monoclonal Antibody (Catalog # MAB7245) followed by HRP-conjugated secondary antibody. A specific band was detected for Rab27a at approximately 26 kDa (as indicated) in the parental U-87 MG cell line, but is significantly reduced in knockdown U-87 MG 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.