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RAB27A+RAB27B Recombinant Rabbit Monoclonal Antibody [PSH24-34]

Usd: 385

Specification

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Catalog# HA724441

RAB27A+RAB27B Recombinant Rabbit Monoclonal Antibody [PSH24-34]

Application

  • WB

  • FC(Intra)

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • 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

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

SwissProt: P51159 Human

SwissProt: O00194 Human

SwissProt: Q9ERI2 Mouse

SwissProt: Q99P58 Mouse

SwissProt: P23640 Rat

SwissProt: Q99P74 Rat

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

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of RAB27A+RAB27B on different lysates with Rabbit anti-RAB27A+RAB27B antibody (<a href="/products/HA724441" style="font-weight: bold;text-decoration: underline;">HA724441</a>) at 1/5,000 dilution.<br /><br />Lane 1: SK-MEL-28 (Human malignant melanoma cells) cell lysate<br />Lane 2: Raji (Human Burkitt's lymphoma cells) cell lysate<br />Lane 3: HL-60 (Human acute promyelocytic leukemia cells) cell lysate<br />Lane 4: Jurkat (Human T-lymphoblastic cells) cell lysate<br />Lane 5: K-562 (Human chronic myelogenous leukemia cells) cell lysate<br />Lane 6: U-87 MG (Human glioblastoma cells) cell lysate<br />Lane 7: NCI-H226 (Human pleural mesothelioma cells) cell lysate<br />Lane 8: PC-12 (Rat pheochromocytoma cells (undifferentiated)) cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br />Exposure time: 10 seconds; ECL: K1801<br /><br />Negative control: Raji (PMID: 15548590).<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/HA724441" style="font-weight: bold;text-decoration: underline;">HA724441</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 25 kDa<br />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: 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

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of RAB27A+RAB27B on different lysates with Rabbit anti-RAB27A+RAB27B antibody (<a href="/products/HA724441" style="font-weight: bold;text-decoration: underline;">HA724441</a>) at 1/5,000 dilution.<br /><br />Lane 1: Mouse bone marrow tissue lysate<br />Lane 2: Mouse lung tissue lysate<br />Lane 3: Mouse skeletal muscle tissue lysate<br />Lane 4: Rat bone marrow tissue lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br />Exposure time: 10 seconds; ECL: K1801<br /><br />Negative control: Mouse skeletal muscle (PMID: 14617806).<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/HA724441" style="font-weight: bold;text-decoration: underline;">HA724441</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 25 kDa<br />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) <br /> <br />Species: Rat <br />Sample: PC-12 (Rat pheochromocytoma cells (undifferentiated)) <br /> <br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. <br />Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature. <br />Antibody dilution buffer: 1x PBS. <br /> <br />Primary antibody: <a href="/products/HA724441" style="font-weight: bold;text-decoration: underline;">HA724441</a>(1/1,000, Red) compared with Rabbit IgG Isotype Control (<a href="/products/HA722127" style="font-weight: bold;text-decoration: underline;">HA722127</a>, Green), 15 minutes at room temperature. <br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 15 minutes at room temperature. <br />

    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) <br /> <br />RAB27A+RAB27B was immunoprecipitated in 0.2 mg K-562 (Human chronic myelogenous leukemia cell) cell lysate with <a href="/products/HA724441" style="font-weight: bold;text-decoration: underline;">HA724441</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA724441" style="font-weight: bold;text-decoration: underline;">HA724441</a> at 1/5,000 dilution. Alpaca anti-Rabbit IgG Fc, Recombinant VHH monoclonal Antibody(<a href="/products/HA1031" style="font-weight: bold;text-decoration: underline;">HA1031</a>) at 1/50000 dilution was used for 1 hour at room temperature. <br /> <br />Lane 1: K-562 cell lysate (input) <br />Lane 2: <a href="/products/HA724441" style="font-weight: bold;text-decoration: underline;">HA724441</a> IP in K-562 cell lysate <br />Lane 3: Rabbit IgG instead of <a href="/products/HA724441" style="font-weight: bold;text-decoration: underline;">HA724441</a> in K-562 cell lysate <br /> <br />Exposure time: 2 seconds <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Primary dilution: <a href="/products/HA724441" style="font-weight: bold;text-decoration: underline;">HA724441</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), 2 hours at room temperature <br /> <br />Predicted band size: 25 kDa <br />Observed band size: 27 kDa

    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

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