Recombinant Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [MJF-R23] to RAB10 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-RAB10 antibody [MJF-R23] - BSA and Azide free
See all RAB10 primary antibodies -
Description
Rabbit monoclonal [MJF-R23] to RAB10 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IF, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: A549, HeLa, HCT 116, MCF7, NIH/3T3, PC-12 and C6 whole cell lysates. ICC/IF: A549 and MCF7 cells. Flow: A549 cells. IP: A549 whole cell lysate.
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General notes
ab238655 is the carrier-free version of ab237703.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
MJF-R23 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab238655 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 22 kDa.
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ICC/IF |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 22 kDa. |
ICC/IF
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
Target
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Function
The small GTPases Rab are key regulators of intracellular membrane trafficking, from the formation of transport vesicles to their fusion with membranes. Rabs cycle between an inactive GDP-bound form and an active GTP-bound form that is able to recruit to membranes different set of downstream effectors directly responsible for vesicle formation, movement, tethering and fusion (By similarity). That Rab is mainly involved in the biosynthetic transport of proteins from the Golgi to the plasma membrane. Regulates, for instance, SLC2A4/GLUT4 glucose transporter-enriched vesicles delivery to the plasma membrane. In parallel, it regulates the transport of TLR4, a toll-like receptor to the plasma membrane and therefore may be important for innate immune response. Plays also a specific role in asymmetric protein transport to the plasma membrane within the polarized neuron and epithelial cells. In neurons, it is involved in axonogenesis through regulation of vesicular membrane trafficking toward the axonal plasma membrane while in epithelial cells, it regulates transport from the Golgi to the basolateral membrane. Moreover, may play a role in the basolateral recycling pathway and in phagosome maturation. According to PubMed:23263280, may play a role in endoplasmic reticulum dynamics and morphology controlling tubulation along microtubules and tubules fusion. -
Sequence similarities
Belongs to the small GTPase superfamily. Rab family. -
Cellular localization
Cytoplasmic vesicle membrane. Golgi apparatus membrane. Golgi apparatus, trans-Golgi network membrane. Endosome membrane. Recycling endosome membrane. Cytoplasmic vesicle, phagosome membrane. Cell projection, cilium. Endoplasmic reticulum membrane. Associates with SLC2A4/GLUT4 storage vesicles (PubMed:22908308). Localizes to the base of the cilium (PubMed:20576682). Transiently associates with phagosomes (By similarity). Localizes to the endoplasmic reticulum at domains of new tubule growth (PubMed:23263280). - Information by UniProt
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Database links
- Entrez Gene: 10890 Human
- Entrez Gene: 19325 Mouse
- Entrez Gene: 50993 Rat
- Omim: 612672 Human
- SwissProt: P61026 Human
- SwissProt: P61027 Mouse
- SwissProt: P35281 Rat
- Unigene: 467960 Human
see all -
Alternative names
- GTP binding protein RAB10 antibody
- Rab10 antibody
- RAB10 member RAS oncogene family antibody
see all
Images
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All lanes : Anti-RAB10 antibody [MJF-R23] (ab237703) at 1/1000 dilution
Lane 1 : Wild-type A549 whole cell lysate
Lane 2 : RAB10 knockout A549 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : MCF7 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 22 kDaThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237703)
Lanes 1 - 4: Merged signal (red and green). Green - ab237703 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab237703 was shown to recognize RAB10 in wild-type A549 cells as signal was lost at the expected MW in RAB10 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and RAB10 knockout samples were subjected to SDS-PAGE. Ab237703 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)Image courtesy of Dr. Dario Alessi from University of Dundee
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237703).
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% saponin permeabilized A549 wild-type and knock-out cells labeling RAB10 (red) with ab237703 at 0.5 μg/ml, followed by anti-Rabbit secondary at 1/1000 dilution. The nuclear counter stain is DAPI (blue).
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All lanes : Anti-RAB10 antibody [MJF-R23] (ab237703) at 1/1000 dilution
Lane 1 : Wild-type A549 (human lung carcinoma epithelial cell line) whole cell lysate
Lane 2 : Rab10 knockout A549 whole cell lysate
Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell line) whole cell lysate
Lane 4 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
Lane 5 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
Lane 6 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 7 : C6 (rat glial tumor glial cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 22 kDa
Observed band size: 22 kDaThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237703)
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
The WT and Rab10 KO A549 lysates were kindly provided by our collaborator Dr. Dario Alessi, University of Dundee.
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Anti-RAB10 antibody [MJF-R23] (ab237703) at 1/10000 dilution + HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 22 kDa
Observed band size: 22 kDa
Exposure time: 180 secondsThis data was developed using ab237703, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
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Immunocytochemistry/ Immunofluorescence - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (human breast adenocarcinoma cell line) cells labeling RAB10 (green) with ab237703 at 1/500 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. Confocal image showing cytoplasmic staining in MCF7 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237703).
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RAB10 was immunoprecipitated from 0.35 mg A549 (human lung carcinoma cell line) whole cell lysate using ab237703 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab237703 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/5000 dilution was used for detection.
Lane 1: A549 whole cell lysate 10μg (input)
Lane 2: ab237703 IP in A549 whole cell lysate.
Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab237703 in A549 whole cell lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237703).
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized A549 (human lung carcinoma cell line) cells labeling RAB10 with ab237703 at 1/600 dilution (red) compared with the rabbit monoclonal IgG (ab172730) isotype control (black) and an unlabelled control (cells without incubation with primary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237703).
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Immunocytochemistry/ Immunofluorescence - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (human lung carcinoma cell line) cells labeling RAB10 (green) with ab237703 at 1/500 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. Confocal image showing cytoplasmic staining in A549 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237703).
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Immunocytochemistry/ Immunofluorescence - Anti-RAB10 antibody [MJF-R23] - BSA and Azide free (ab238655)Image courtesy of Dr. Dario Alessi from University of Dundee
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab237703).
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% saponin permeabilized Human A549 wild-type and knock-out cells labeling RAB10 (red) with ab237703 at 0.5 μg/ml, followed by anti-Rabbit secondary at 1/1000 dilution. The nuclear counter stain is DAPI (blue).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab238655 has not yet been referenced specifically in any publications.