Anti-Rab5 antibody - Early Endosome Marker (ab18211)Anti-Rab5早期核內(nèi)體標(biāo)記產(chǎn)品信息:
Product nameAnti-Rab5 antibody - Early Endosome MarkerSee all Rab5 primary antibodies ...
Description
Rabbit polyclonal to Rab5 - Early Endosome Marker
Tested applicationsIP, IHC-FrFl, IHC-Fr, ICC/IF, WB more details
Species reactivity
Reacts 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.
Read Abcam's proprietary immunogen policy
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
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.
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 approximay 24 kDa (predicted molecular weight: 24 kDa).Can be blocked with Human 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.
Information by UniProt
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
Alternative names
RAB 5 antibody
RAB 5A antibody
RAB5A antibody
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 Pezet
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.
Immunoprecipitation - Rab5 antibody - Early Endosome Marker (ab18211)This image is courtesy of an anonymous Abreview
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.
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 + Human Rab5 full length protein (ab62956) at 0.001 µg Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution developed using the ECL technique Performed under reducing conditions. Exposure time : 150 seconds