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Immunogen
Human Rab5 synthetic peptide conjugated to KLH; identical to dog Rab5 sequence over the residues
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Antibody s target
Human Rab5
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Antibody s full description
Rabbit Anti-Human Rab5 Polyclonal
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Primary research fields
Cell Signaling, Organelle Markers, Trafficking, Neuroscience, Cell Structure, Pre-Synaptic Markers
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Antibody s category
Polyclonal Antibodies
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Antibody s other name
Rab 5A Antibody, RAS associated protein RAB5A Antibody, Ras related protein Rab 5 A Antibody
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Verified applications
WB, IHC, ICC/IF
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Raised in
Rabbit
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Antibody s reactivity
Human, Mouse, Rat, Bovine, Monkey
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Antibody s recommended dilutions for use
WB (1:1000), IHC (1:100), ICC/IF (1:80); optimal dilutions for assays should be determined by the user.
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Antibody s purified from
Protein A Purified
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Recommended buffer for storage
PBS, 50% glycerol, 0.09% sodium azide
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Antibody s concentration
1 mg/ml
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Antibody s specificity
Detects ~26kDa.
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Storage recommendations
-20°C
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Shipping recommendations
Blue Ice or 4°C
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Certificate of analysis
1 µl/ml of SPC-168 was sufficient for detection of Rab5 in 15 µg of HeLa lysate by ECL immunoblot analysis using Donkey anti-rabbit IgG:HRP as the secondary antibody.
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Antibody in cell
Cell Membrane, Endosome, Early Endosome Membrane, Melanosome
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Tissue specificity
See included datasheet or contact our support service.
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Scientific context
Rab5 is a 24kDa member of the Rab family of small guanosine triphosphatases (GTPases), Ras superfamily. Rab GTPases are central regulators of membrane trafficking in the eukaryotic cell. Their regulatory capacity depends on their ability to cycle between the GDP -bound inactive and GTP-bound active states. This conversion is regulated by GDP/GTP exchange factors (GEPs), GDP dissociation inhibitors (GDIs) and GTPase-activating proteins (GAPs) (1, 2). Activation of a Rab protein is coupled to its association with intracellular membranes, allowing it to recruit downstream effector proteins to the cytoplasmic surface of a subcellular compartment (3). Through these proteins, Rab GTPases regulate vesicle formation, actin- and tubulin-dependent vesicle movement, and membrane fusion(1). Rab proteins contain conserved regions involved in guanine-nucleotide binding, and hyper variable COHO-terminal domains with a cysteine motif implicated in subcellular targeting. Post-translational modification of the cysteine motif with one or two geranyl groups is essential for the membrane association and correct intracellular localization of Rab proteins(3). Each Rab shows a characteristic subcellular distribution (4). In particular, Rab5 is ubiquitously expressed in human tissues. It localizes mainly to early endosomes, but is also present on the plasma membrane. It regulates the fusion between endocytic vesicles and early endosomes, as well as the homotypic fusion between early endosomes (5). Among the proteins recruited by the GTP-bound active Rab5 are Rabaptin-5 and EEA1 (6). Anti-Rab5 may be used as an early endosome marker.
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Bibliography
1. Stenmark H., and Olkkonen V.M. (2001) Genome Biol. 2: 3007.1-3007.7. 2. Takai Y., et al. (2001) Physiol. Rev. 8:, 153-208. 3. Ali B.R., et al. (2004) J. Cell Sci. 117: 6401-6412. 4. Zerial M., and McBride H. (2001) Nat. Rev. Mol. Cell Biol. 2: 107-117. 5. Sonnichsen B., et al. (2000) J. Cell Biol. 149: 901-913 6. Woodman P.G. (2000) Traffic. 1: 695-701.
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Released date
13/Aug/2009
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NCBI number
NP_004153.2
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Gene number
5868
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Protein number
Q6FI44
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PubMed number
21586748
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Tested applications
Immunocytochemistry/Immunofluorescence
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Tested reactivity
Human
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Antibody s datasheet
Contact our support service to receive datasheet or other technical documentation.
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Representative figure link
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Representative figure legend
Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-Rab5 Polyclonal Antibody (SPC-168). Tissue: HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rabbit Anti-Rab5 Polyclonal Antibody (SPC-168) at 1:80 for 12 hours at 4°C. Secondary Antibody: R-PE Goat Anti-Rabbit (yellow) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Melanosome. Nucleus. Magnification: 100x. (A) DAPI (blue) nuclear stain. (B) Anti-Rab5 Antibody. (C) Composite. | Western blot analysis of Human Cell line lysates showing detection of Rab5 protein using Rabbit Anti-Rab5 Polyclonal Antibody (SPC-168). Load: 15 µg protein. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Rabbit Anti-Rab5 Polyclonal Antibody (SPC-168) at 1:1000 for 2 hours at RT. Secondary Antibody: Donkey Anti-Rabbit IgG: HRP for 1 hour at RT. | Immunohistochemistry analysis using Rabbit Anti-Rab5 Polyclonal Antibody (SPC-168). Tissue: backskin. Species: Mouse. Fixation: Bouin's Fixative Solution. Primary Antibody: Rabbit Anti-Rab5 Polyclonal Antibody (SPC-168) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Rabbit (green) at 1:50 for 1 hour at RT. Localization: Cytoplasm. | Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-Rab5 Polyclonal Antibody (SPC-168). Tissue: HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rabbit Anti-Rab5 Polyclonal Antibody (SPC-168) at 1:80 for 12 hours at 4°C. Secondary Antibody: FITC Goat Anti-Rabbit (green) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Melanosome. Nucleus. Magnification: 20x. (A) DAPI (blue) nuclear stain. (B) Anti-Rab5 Antibody. (C) Composite. Rabbit Anti-Rab5 Antibody used in Immunocytochemistry/Immunofluorescence (ICC/IF) on Human HeLa Cells (SPC-168) | Rabbit Anti-Rab5 Antibody used in Western blot (WB) on Human Cell line lysates (SPC-168) | Rabbit Anti-Rab5 Antibody used in Immunohistochemistry (IHC) on Mouse backskin (SPC-168) | Rabbit Anti-Rab5 Antibody used in Immunocytochemistry/Immunofluorescence (ICC/IF) on Human HeLa Cells (SPC-168)
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Warning information
Non-hazardous
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Country of production
Canada
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Total weight kg
1.4
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Net weight g
0.1
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Stock availabilit
In Stock
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Description
This antibody needs to be stored at + 4°C in a fridge short term in a concentrated dilution. Freeze thaw will destroy a percentage in every cycle and should be avoided. Antibody for research use.
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Test
StressMark antibodies supplies antibodies that are for research of human proteins.
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Properties
Human proteins, cDNA and human recombinants are used in human reactive ELISA kits and to produce anti-human mono and polyclonal antibodies. Modern humans (Homo sapiens, primarily ssp. Homo sapiens sapiens). Depending on the epitopes used human ELISA kits can be cross reactive to many other species. Mainly analyzed are human serum, plasma, urine, saliva, human cell culture supernatants and biological samples.
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Group
Polyclonals and antibodies
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About
Polyclonals can be used for Western blot, immunohistochemistry on frozen slices or parrafin fixed tissues. The advantage is that there are more epitopes available in a polyclonal antiserum to detect the proteins than in monoclonal sera. Rabbits are used for polyclonal antibody production by StressMark antibodies. Rabbit antibodies are very stable and can be stored for several days at room temperature. StressMark antibodies adds sodium azide and glycerol to enhance the stability of the rabbit polyclonal antibodies. Anti-human, anti mouse antibodies to highly immunogenic selected peptide sequences are" monoclonal like" since the epitope to which they are directed is less than 35 amino acids long.
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Latin name
Oryctolagus cuniculus