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Western blot - Rab5 antibody (ab18211)
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Immunocytochemistry/ Immunofluorescence - Rab5 antibody - Early Endosome Marker (ab18211)
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Western blot - Rab5 antibody - Early Endosome Marker (ab18211)
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Product Name
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Rab5 antibody - Early Endosome Marker
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See all Rab5 antibodies (9)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to Rab5 - Early Endosome Marker
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Immunogen
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Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human Rab5.
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Reacts with
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Hu, Ms, Rat
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Predicted to react
(species key)
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Dog
(Due to sequence homology)
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Tested applications
(see key)
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ICC/IF, IHC-Fr, IHC-FrFl, IP, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ICC/IF: Use at a concentration of 1 µg/ml.
IHC-Fr: 1/300.
IHC-FrFl: Use at an assay dependent concentration.
IP: Use at an assay dependent concentration (see abreview).
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 24 kDa (predicted molecular weight: 24 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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This antibody gave a positive control in the following human lysate:
Jurkat (Human T cell lymphoblast-like cell line) whole cell
This antibody gave a positive control in the following mouse lysates:
NIH 3T3 whole cell
MEF1 whole cell
Brain tissue
Testis tissue
Spinal Cord tissue
This antibody gave a positive control in the following rat lysates:
PC12 whole cell
Brain tissue
Heart tissue
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Cellular localization
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Cell membrane; Lipid-anchor; Cytoplasmic side. Early endosome membrane; Lipid-anchor. Melanosome.
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Research areas
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Neuroscience >> Cell Type Marker >> Neuron marker >> Synapse marker
Signal Transduction >> Protein Trafficking >> Vesicle Transport >> Regulation
Tags & Cell Markers >> Subcellular Markers >> Organelles >> Endosome
Neuroscience >> Neurotransmission >> Secretory Vesicles >> Rabs
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Relevance
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Rab5 is a 24 kDa GTP-binding protein that regulates the fusion of plasma membrane-derived clathrin-coated vesicles with early endosomes and homotypic fusion among early endosomes. It is localized to the cytoplasmic side of the plasma membrane, clathrin coated vesicles and early endosomes. Rab5 is believed to regulate vesicle fusion through a cycle of GDP/GTP exchange and GTP hydrolysis. The different guanine nucleotide binding states of Rab5 is postulated to affect its ability to associate or dissociate with membranes during endocytotic membrane traffic. Its GTP-bound form, which represents the active form of Rab5, associates with membrane and regulates vesicle docking and fusion. Studies using a Rab5 mutant that hydrolysed xanthosine 5'-triphosphate (XTP) indicated that nucleotide hydrolysis occurs even in the absence of membrane fusion. GTP hydrolysis by Rab5 is postulated to determine the frequency of membrane docking and fusion events.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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Batch dependent within range: 0.10 - 0.20 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this Rab5 antibody - Early Endosome Marker is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for Rab5 antibody - Early Endosome Marker images, references, products related to ab18211 and other tools.
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Rab5 antibody - Early Endosome Marker images:
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 Western blot - Rab5 antibody (ab18211)
Rab5 antibody - Early Endosome Marker (ab18211) at 1 µg/ml + Jurkat cell lysate at 20 µg
Secondary
Alexa Fluor Goat polyclonal to Rabbit IgG (700) at 1/5000 dilution
Predicted band size : 24 kDa
Observed band size : 24 kDa
Additional bands at : 40 kDa,70 kDa. We are unsure as to the identity of these extra bands.
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Immunocytochemistry/ Immunofluorescence - Rab5 antibody - Early Endosome Marker (ab18211)
ICC/IF image of ab18211 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab18211, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
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Western blot - Rab5 antibody - Early Endosome Marker (ab18211)
All lanes : Rab5 antibody - Early Endosome Marker (ab18211) at 1 µg/ml
Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)
Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab46770)
Lane 3 : Brain (Mouse) Tissue Lysate (ab27253)
Lane 4 : Testis (Mouse) Tissue Lysate - normal tissue
Lane 5 : Spinal Cord (Mouse) Tissue Lysate
Lane 6 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lane 7 : Brain (Rat) Tissue Lysate - normal tissue
Lane 8 : Heart (Rat) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 24 kDa
Observed band size : 25 kDa (why is the actual band size different from the predicted?)
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Immunohistochemistry (Frozen sections) - Rab5 antibody - Early Endosome Marker (ab18211)
ab18211 at 1/300 staining adult rat brain (perfusion fixed) tissue sections by IHC-Fr. Adult rat was perfused intracardially with paraformaldehyde 4% in PB 0.2M. The brain was post-fixed in the same fixative for 24 hours. Sections were cryoprotected with sucrose 20% and later frozen in OCT. Sections were incubated in free floating for 12h with the primary antibody (1/300) and later revealed with secondary antibody conjugated with Alexa Fluor ® 488 (1/2000). The staining obtained is restricted to the cytoplasm and consists of a small and thin punctate staining. The picture shows the staining obtained at the level of the spinal cord using the X20 objective and zooming on two particular neurons. This image is courtesy of an Abreview submitted by Dr Sophie Pezet
See Abreview
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Immunoprecipitation - Rab5 antibody - Early Endosome Marker (ab18211)
ab18211 immunoprecipitated Rab5 in Mouse neuroblastoma N2a whole cell lysate. 10µg of cell lysate was incubaed with primary antibody (1/2000 in dilution buffer:0.025 M Tris, 0.15 M NaCl, 0.001 M EDTA, 1% NP-40, 5% glycerol pH 7.4) for 2 hours at 22°C and an AminoLink® Plus Coupling Resin matrix.
For Western blotting an HRP conjugated goat monoclonal to rabbit IgG (1/2000) was used. This image is courtesy of an anonymous Abreview
See Abreview
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References for Rab5 antibody - Early Endosome Marker (ab18211)
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This product has been used in: (two most recent references)
Frykman S et al. Synaptic and endosomal localization of active gamma-secretase in rat brain. PLoS One 5:e8948 (2010). ICC/IF, IHC-Fr; Mouse, Rat. PubMed: 20126630
Hooper S et al. A chemical biology screen reveals a role for Rab21-mediated control of actomyosin contractility in fibroblast-driven cancer invasion. Br J Cancer 102:392-402 (2010). WB, ICC/IF; Human. PubMed: 19953096
See all 4 publication references for this product.
If you publish research using ab18211 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to Rab5
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Rab5 antibody - Early Endosome Marker - more information
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