| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, FC(Intra), IP |
| Clonality: | Monoclonal |
| Clone number: | PSH24-34 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage buffer: | 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 25 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human RAB27A aa 1-221. |
| Positive control: | SK-MEL-28 (Human malignant melanoma cells) cell lysate, HL-60 (Human acute promyelocytic leukemia cells) cell lysate, Jurkat (Human T-lymphoblastic cells) cell lysate, K-562 (Human chronic myelogenous leukemia cells) cell lysate, U-87 MG (Human glioblastoma cells) cell lysate, NCI-H226 (Human pleural mesothelioma cells) cell lysate, PC-12 (Rat pheochromocytoma cells (undifferentiated)) cell lysate, Mouse bone marrow tissue lysate, Mouse lung tissue lysate, Rat bone marrow tissue lysate. |
| Subcellular location: | Membrane, Melanosome, Late endosome, Lysosome. |
| Uniprot #: | SwissProt: P51159 Human | O00194 Human | Q9ERI2 Mouse | Q99P58 Mouse | P23640 Rat | Q99P74 Rat |
| Alternative names: | Ras-related protein Rab-27A EC:3.6.5.2 Rab-27 GTP-binding protein Ram RAB27A RAB27 Ras-related protein Rab-27B C25KG RAB27B |
|
Fig1:
Western blot analysis of RAB27A+RAB27B on different lysates with Rabbit anti-RAB27A+RAB27B antibody (HA724441) at 1/5,000 dilution. Lane 1: SK-MEL-28 (Human malignant melanoma cells) cell lysate Lane 2: Raji (Human Burkitt's lymphoma cells) cell lysate Lane 3: HL-60 (Human acute promyelocytic leukemia cells) cell lysate Lane 4: Jurkat (Human T-lymphoblastic cells) cell lysate Lane 5: K-562 (Human chronic myelogenous leukemia cells) cell lysate Lane 6: U-87 MG (Human glioblastoma cells) cell lysate Lane 7: NCI-H226 (Human pleural mesothelioma cells) cell lysate Lane 8: PC-12 (Rat pheochromocytoma cells (undifferentiated)) cell lysate Lysates/proteins at 15 µg/Lane. Exposure time: 10 seconds; ECL: K1801 Negative control: Raji (PMID: 15548590). Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA724441, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 25 kDa Observed band size: 27 kDa |
|
Fig2:
Western blot analysis of RAB27A+RAB27B on different lysates with Rabbit anti-RAB27A+RAB27B antibody (HA724441) at 1/5,000 dilution. Lane 1: Mouse bone marrow tissue lysate Lane 2: Mouse lung tissue lysate Lane 3: Mouse skeletal muscle tissue lysate Lane 4: Rat bone marrow tissue lysate Lysates/proteins at 15 µg/Lane. Exposure time: 10 seconds; ECL: K1801 Negative control: Mouse skeletal muscle (PMID: 14617806). Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA724441, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 25 kDa Observed band size: 27 kDa |
|
Fig3:
Application: Flow Cytometry (Intra)
Species: Rat Sample: PC-12 (Rat pheochromocytoma cells (undifferentiated)) Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature. Antibody dilution buffer: 1x PBS. Primary antibody: HA724441(1/1,000, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature. Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature. |
|
Fig4:
Immunoprecipitation (IP)
RAB27A+RAB27B was immunoprecipitated in 0.2 mg K-562 (Human chronic myelogenous leukemia cell) cell lysate with HA724441 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724441 at 1/5,000 dilution. Alpaca anti-Rabbit IgG Fc, Recombinant VHH monoclonal Antibody(HA1031) at 1/50000 dilution was used for 1 hour at room temperature. Lane 1: K-562 cell lysate (input) Lane 2: HA724441 IP in K-562 cell lysate Lane 3: Rabbit IgG instead of HA724441 in K-562 cell lysate Exposure time: 2 seconds Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary dilution: HA724441, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature Predicted band size: 25 kDa Observed band size: 27 kDa |