RAB27A+RAB27B Recombinant Rabbit Monoclonal Antibody [PSH24-34]
cat.: HA724441
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
Images
HA724441_1.jpg 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
HA724441_2.jpg 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
HA724441_3.jpg 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.
HA724441_4.jpg 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
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.