Anti-Rab5 antibody - Early Endosome Marker (ab18211)
- Datasheet
- Specific References (52)
- Protocols
Overview
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Product nameAnti-Rab5 antibody - Early Endosome Marker
See all Rab5 primary antibodies -
DescriptionRabbit polyclonal to Rab5 - Early Endosome Marker
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Host speciesRabbit
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SpecificityThe immunogen used for this product shares 43% homology with Rab5C. Cross-reactivity with this protein has not been confirmed experimentally.
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Tested applicationsSuitable for: IP, IHC-FrFl, IHC-Fr, ICC/IF, WBmore details
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Species reactivityReacts with: Mouse, Rat, Dog, Human, Chinese hamster
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human Rab5.
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Positive control
- This antibody gave a positive control in the following human lysate: Jurkat (Human T cell lymphoblast-like cell line) whole cell, NIH 3T3 whole cell, MEF1 whole cell, Brain tissue, Testis tissue, Spinal Cord tissue, PC12 whole cell, Brain tissue and Heart tissue ICC/IF: HepG2
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Positive Controls
Applications
Our Abpromise guarantee covers the use of ab18211 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 |
|---|---|---|
| IP | Use at an assay dependent concentration. | |
| IHC-FrFl | Use at an assay dependent concentration. | |
| IHC-Fr | 1/300. | |
| ICC/IF | Use a concentration of 1 - 5 µg/ml. we recommend to perform a methanol fixation on the cells |
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| WB | Use a concentration of 1 µg/ml. Detects a band of approximately 24 kDa (predicted molecular weight: 24 kDa). |
Target
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FunctionRequired for the fusion of plasma membranes and early endosomes.
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Sequence similaritiesBelongs to the small GTPase superfamily. Rab family.
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Cellular localizationCell membrane. Early endosome membrane. Melanosome. Enriched in stage I melanosomes.
- Information by UniProt
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Database links
- Entrez Gene: 404008 Dog
- Entrez Gene: 5868 Human
- Entrez Gene: 271457 Mouse
- Entrez Gene: 64633 Rat
- Omim: 179512 Human
- SwissProt: P18066 Dog
- SwissProt: P20339 Human
- SwissProt: Q9CQD1 Mouse
see all -
Alternative names
- RAB 5 antibody
- RAB 5A antibody
- RAB5A antibody
see all
Images
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Western blot - Anti-Rab5 antibody - Early Endosome Marker (ab18211)Anti-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. -
Immunocytochemistry/ Immunofluorescence - Anti-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 - Anti-Rab5 antibody - Early Endosome Marker (ab18211)All lanes : Anti-Rab5 antibody - Early Endosome Marker (ab18211) at 1 µg/ml
Lane 1 :NIH 3T3 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
All lanes : 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?) -
Immunohistochemistry (Frozen sections) - Anti-Rab5 antibody - Early Endosome Marker (ab18211)This image is courtesy of an Abreview submitted by Dr Sophie Pezetab18211 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.
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Immunoprecipitation - Anti-Rab5 antibody - Early Endosome Marker (ab18211)This image is courtesy of an anonymous Abreviewab18211 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. -
Western blot - Anti-Rab5 antibody - Early Endosome Marker (ab18211)
Protocols
References
This product has been referenced in:
- Panigrahi GK et al. Hypoxia-induced exosome secretion promotes survival of African-American and Caucasian prostate cancer cells. Sci Rep 8:3853 (2018). Read more (PubMed: 29497081) »
- Sun Y et al. SFTS phlebovirus promotes LC3-II accumulation and nonstructural protein of SFTS phlebovirus co-localizes with autophagy proteins. Sci Rep 8:5287 (2018). Read more (PubMed: 29588492) »
Customer reviews and Q&As
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"