RAB27A+RAB27B Recombinant Rabbit Monoclonal Antibody [PSH24-34]
Catalog# HA724441
RAB27A+RAB27B Recombinant Rabbit Monoclonal Antibody [PSH24-34]
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WB
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FC(Intra)
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IP
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
RAB27A+RAB27B Recombinant Rabbit Monoclonal Antibody [PSH24-34]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within human RAB27A aa 1-221.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, FC(Intra), IP
Target Molecular Weight
Predicted band size: 25 kDa
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.
Conjugation
unconjugated
Clone Number
PSH24-34
Reactivity Data
Verified (internally validated)
Reported in literature (not internally validated)
Predicted reactive (based on sequence homology)
Not recommended (failed internal validation)
| WB | IHC-P | FC(Intra) | IF-Cell | IP | |
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| mouse |
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| rat |
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Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
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.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
Target
Function
Rab27 is a member of the Rab subfamily of GTPases. Rab27 is post-translationally modified by the addition of two geranylgeranyl groups on the two C-terminal cysteines. Rab27 has two main isoforms: Rab27a and Rab27b. These are similar in primary composition, with 66% similarity between nucleotides. Most of their differences originate from their C-terminal, which is responsible for interactions with proteins such as SPIREs. Thus, these isoforms play different roles in the regulation pathway of exocytosis. Throughout the process of exocytosis, Rab27a and Rab27b are found in different sections of the cell, with Rab27b found commonly in the TGN and Rab27a usually bound to multivesicular endosomes with CD63 present. Rab27 plays a key role in the regulation of exocytosis of vesicles in various cellular organelles. In certain secretory organelles, such as Weibel-Palade bodies in endothelial cells, Rab27a is recruited during organelle maturation, rendering them from a secretion-incompetent to a secretion-comptetent state. Rab27 uses effectors to tether vesicles to the cytoskeleton and transport them to the plasma membrane, where they undergo fusion. They ensure that vesicles attach correctly, in the proper orientation, at the dedicated site of fusion. However, fusion itself is started when effectors bind SNARE proteins that catalyze the start of exocytosis.
Background References
1. An HJ et al. RAB27A and RAB27B Expression May Predict Lymph Node Metastasis and Survival in Patients With Gastric Cancer. Cancer Genomics Proteomics. 2022 Sep-Oct
Subcellular Location
Membrane, Melanosome, Late endosome, Lysosome.
UNIPROT
Synonyms
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
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☑ Relative expression (RE)
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 -
☑ Relative expression (RE)
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 -
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. -
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
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"