Product datasheet
Anti-Rab5 antibody - Early Endosome Marker ab18211
22 Abreviews 20 References 6 Images
Overview
Product name
Anti-Rab5 antibody - Early Endosome Marker
Description
Rabbit polyclonal to Rab5 - Early Endosome Marker
Tested applications
IP, IHC-FrFl, IHC-Fr, ICC/IF, WB
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 ICC/IF: HepG2
Properties
Form
Liquid
Storage instructions
Shipped 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 buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
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.
1
Application
Abreviews
Notes
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).
Target
Function
Required for the fusion of plasma membranes and early endosomes.
Sequence similarities
Belongs to the small GTPase superfamily. Rab family.
Cellular localization
Cell membrane. Early endosome membrane. Melanosome. Enriched in stage I melanosomes.
Anti-Rab5 antibody - Early Endosome Marker images
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
Western blot - Rab5 antibody (ab18211)
are unsure as to the identity of these extra
bands.
2
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 TBST) was used for all other blocking steps. DAPI
was used to stain the cell nuclei (blue). Alexa
Immunocytochemistry/ Immunofluorescence -
Fluor® 594 WGA was used to label plasma
Rab5 antibody - Early Endosome Marker
membranes (red).
(ab18211)
All lanes : Anti-Rab5 antibody - Early
Endosome Marker (ab18211) at 1 µg/ml
Lane 1 : NIH 3T3 (Mouse) 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
Western blot - Rab5 antibody - Early Endosome
Lane 5 : Spinal Cord (Mouse) Tissue Lysate
Marker (ab18211)
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
3
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
Immunohistochemistry (Frozen sections) - Rab5
later revealed with secondary antibody
antibody - Early Endosome Marker (ab18211)
conjugated with Alexa Fluor ® 488 (1/2000).
This image is courtesy of an Abreview submitted by
Dr Sophie Pezet
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% NP40, 5% glycerol pH 7.4) for 2 hours at 22°C
and an AminoLink® Plus Coupling Resin
Immunoprecipitation - Rab5 antibody - Early
matrix.
Endosome Marker (ab18211)
For Western blotting an HRP conjugated goat
This image is courtesy of an anonymous Abreview
monoclonal to rabbit IgG (1/2000) was used.
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 24 kDa
Exposure time : 150 seconds
Western blot - Anti-Rab5 antibody - Early
Endosome Marker (ab18211)
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
4
Our Abpromise to you: Quality guaranteed and expert technical support
Replacement or refund for products not performing as stated on the datasheet
Valid for 12 months from date of delivery
Response to your inquiry within 24 hours
We provide support in Chinese, English, French, German, Japanese and Spanish
Extensive multi-media technical resources to help you
We investigate all quality concerns to ensure our products perform to the highest standards
If the product does not perform as described on this datasheet, we will offer a refund or replacement. For full details of the Abpromise,
please visit http://www.abcam.com/abpromise or contact our technical team.
Terms and conditions
Guarantee only valid for products bought direct from Abcam or one of our authorized distributors
5