Anti-Rab5 antibody - Early Endosome Marker (ab18211)
Overview
- Product nameAnti-Rab5 antibody - Early Endosome MarkerSee all Rab5 primary antibodies ...
- DescriptionRabbit polyclonal to Rab5 - Early Endosome Marker
- Tested applicationsIP, IHC-FrFl, IHC-Fr, ICC/IF, WB more details
- Species reactivityReacts with: Mouse, Rat, Dog, Human, Chinese Hamster
- Immunogen
Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human Rab5.
- Positive controlThis 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
Properties
- FormLiquid
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... - PurityImmunogen affinity purified
- Clonality Polyclonal
- IsotypeIgG
- Research Areas
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 | Notes |
|---|---|
| IP | IP: Use at an assay dependent concentration. |
| IHC-FrFl | IHC-FrFl: Use at an assay dependent concentration. |
| IHC-Fr | IHC-Fr: 1/300. |
| ICC/IF | ICC/IF: Use a concentration of 1 µg/ml. |
| WB | WB: Use a concentration of 1 µg/ml. Detects a band of approximately 24 kDa (predicted molecular weight: 24 kDa).Can be blocked with Rab5 peptide (ab18625). |
Target
- FunctionRequired for the fusion of plasma membranes and early endosomes.
- Sequence similaritiesBelongs to the small GTPase superfamily. Rab family.
- Cellular localizationCell membrane. Early endosome membrane. Melanosome. Enriched in stage I melanosomes.
-
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
- Unigene: 475663 Human
- Unigene: 329123 Mouse
- Unigene: 44477 Rat
see all
Target information above from: UniProt accession
P20339
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
.
-
Alternative names
- RAB 5 antibodyRAB 5A antibodyRAB5A antibody
- RAB5A member RAS oncogene family antibodyRAB5A_HUMAN antibodyRAS associated protein RAB5A antibodyRas related protein Rab 5A antibodyRas-related protein Rab-5A antibody
see all
Anti-Rab5 antibody - Early Endosome Marker images
-
Western blot - Rab5 antibody (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 - 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).
-
Western blot - Rab5 antibody - Early Endosome Marker (ab18211)All lanes : Anti-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?) -
Immunohistochemistry (Frozen sections) - 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.
-
Immunoprecipitation - 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. -
Immunocytochemistry/ Immunofluorescence - Rab5 antibody - Early Endosome Marker (ab18211)ICC/IF image of ab18211 stained Hela cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab18211, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. -
Western blot - Anti-Rab5 antibody - Early Endosome Marker (ab18211)Anti-Rab5 antibody - Early Endosome Marker (ab18211) at 1 µg/ml +Rab5 protein (ab62956) at 0.001 µg
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Exposure time : 150 seconds
References for Anti-Rab5 antibody - Early Endosome Marker (ab18211)
This product has been referenced in:
- Skorupa A et al. Motoneurons secrete angiogenin to induce RNA cleavage in astroglia. J Neurosci 32:5024-38 (2012). WB ; Mouse . Read more (PubMed: 22496549) »
- Wolf M et al. Rhesus rotavirus entry into a polarized epithelium is endocytosis dependent and involves sequential VP4 conformational changes. J Virol 85:2492-503 (2011). ICC/IF ; Dog . Read more (PubMed: 21191022) »
