RAB27A Recombinant Rabbit Monoclonal Antibody [PSH24-10]
cat.: HA751925
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH24-10 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4). |
| 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, Rat lung tissue lysate. |
| Subcellular location: | Membrane, Melanosome, Late endosome, Lysosome. |
| Recommended Dilutions:
WB IHC-P FC IP |
1:5,000 1:200 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: P51159 Human | Q9ERI2 Mouse | P23640 Rat |
| Alternative names: | GS2 GTP-binding protein Ram HsT18676 MGC117246 Mutant Ras related protein Rab-27A Rab-27 RAB-27A RAB27 RAB27A RAB27A member RAS oncogene family RAM Ras-related protein Rab-27A Ras-related protein Rab27A RB27A_HUMAN |
Images
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Fig1:
Western blot analysis of RAB27A on different lysates with Rabbit anti-RAB27A antibody (HA751925) 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: HA751925, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C 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 |
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Fig2:
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: HA751925 (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. |
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Fig3:
Western blot analysis of RAB27A on different lysates with Rabbit anti-RAB27A antibody (HA751925) 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 Lane 5: Rat lung 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: HA751925, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C 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 |
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Fig4:
Immunoprecipitation (IP)
RAB27A was immunoprecipitated in 0.2 mg K-562 (Human chronic myelogenous leukemia cells) cell lysate with HA751925 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751925 at 1/5,000 dilution. Alpaca anti-Rabbit IgG Fc, Recombinant VHH monoclonal Antibody(HA1031) at 1/50,000 dilution was used for 1 hour at room temperature. Lane 1: K-562 cell lysate (input) Lane 2: HA751925 IP in K-562 cell lysate Lane 3: Rabbit IgG instead of HA751925 in K-562 cell lysate Exposure time: 14 seconds Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary dilution: HA751925, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature Predicted band size: 25 kDa Observed band size: 27 kDa |
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Fig5:
Application: Immunohistochemistry (IHC-P)
Species: Human Tissue: Malignant melanoma Sample: Paraffin-embedded section Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. Wash buffer: 1× TBST Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. Primary antibody: HA751925, 1/200, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. |
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Fig6:
Application: Immunohistochemistry (IHC-P)
Species: Human Tissue: Colon Sample: Paraffin-embedded section Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. Wash buffer: 1× TBST Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. Primary antibody: HA751925, 1/200, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. |
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Fig7:
Application: Immunohistochemistry (IHC-P)
Species: Human Tissue: Brain Sample: Paraffin-embedded section Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. Wash buffer: 1× TBST Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. Primary antibody: HA751925, 1/200, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. Negative expression of RAB27A protein in brain is consistent with the predicted expression pattern. |
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Fig8:
Application: Immunohistochemistry (IHC-P)
Species: Mouse Tissue: Colon Sample: Paraffin-embedded section Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. Wash buffer: 1× TBST Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. Primary antibody: HA751925, 1/200, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. |
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Fig9:
Application: Immunohistochemistry (IHC-P)
Species: Mouse Tissue: Brain Sample: Paraffin-embedded section Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. Wash buffer: 1× TBST Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. Primary antibody: HA751925, 1/200, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. Negative expression of RAB27A protein in brain is consistent with the predicted expression pattern. |
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Fig10:
Application: Immunohistochemistry (IHC-P)
Species: Rat Tissue: Colon Sample: Paraffin-embedded section Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. Wash buffer: 1× TBST Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. Primary antibody: HA751925, 1/200, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. |
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Fig11:
Application: Immunohistochemistry (IHC-P)
Species: Rat Tissue: Brain Sample: Paraffin-embedded section Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. Wash buffer: 1× TBST Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. Primary antibody: HA751925, 1/200, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. Negative expression of RAB27A protein in brain is consistent with the predicted expression pattern. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.