Product nameAnti-Rab5 antibody - Early Endosome Marker
See all Rab5 primary antibodies
DescriptionRabbit polyclonal to Rab5 - Early Endosome Marker
SpecificityThe immunogen used for this product shares 43% homology with Rab5C. Cross-reactivity with this protein has not been confirmed experimentally.
Tested applicationsSuitable for: IP, IHC-FrFl, IHC-Fr, ICC/IF, WBmore details
Species reactivityReacts with: Mouse, Rat, Dog, Human, Chinese hamster
Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human Rab5.
- Recombinant Human Rab5 protein (ab62956) can be used as a positive control in WB. 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
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.
Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
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.
|IP||Use at an assay dependent concentration.|
|IHC-FrFl||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 1 - 5 µg/ml.
we recommend to perform a methanol fixation on the cells
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 24 kDa (predicted molecular weight: 24 kDa).|
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.
- Information by UniProt
- RAB 5 antibody
- RAB 5A antibody
- RAB5A antibody
Anti-Rab5 antibody - Early Endosome Marker (ab18211) at 1 µg/ml + Jurkat cell lysate at 20 µg
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.
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).
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.
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?
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.
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 product has been referenced in:
- Villella VR et al. A pathogenic role for cystic fibrosis transmembrane conductance regulator in celiac disease. EMBO J 38:N/A (2019). Read more (PubMed: 30498130) »
- Moya-Alvarado G et al. Brain-Derived Neurotrophic Factor (BDNF) Regulates Rab5-Positive Early Endosomes in Hippocampal Neurons to Induce Dendritic Branching. Front Cell Neurosci 12:493 (2018). Read more (PubMed: 30618640) »