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Anti-Rab5 antibody - Early Endosome Marker (ab18211)Anti-Rab5早期核内体标记产品信息:
Rabbit polyclonal to Rab5 - Early Endosome Marker
Reacts with: Mouse, Rat, Dog, Human, Chinese Hamster
Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human Rab5.
Read Abcam's proprietary immunogen policy
Properties
Concentration 100 µg at 1 mg/ml
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 approximay 24 kDa (predicted molecular weight: 24 kDa).Can be blocked with Human Rab5 peptide (ab18625). |
Target
Database links
Alternative names
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.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
公司介绍:
Abcam位于英国的剑桥科学园,成立于1998年,专门生产和分销研究型抗体。主要创立人Jonathan Milner和实验室同事构思成立一间了解科研人员需要的网上抗体公司,理念是Abcam销售质量高的抗体,并提供zui全面、诚实、时新的说明书,快速的送货服务,及高效率的顾客和服务。我们的在线目录(www.jiefang996.com) 已有差不多100,000种抗体和试剂,并不断添加,供应予百多个国家。Abcam于2005年11月在伦敦证券交易所上市,在美国马萨诸塞州、日本东京及香港均设有分公司。
Abcam 一直致力加强产品线,为使研究员更容易找到蛋白质研究试剂产品,在2011并购了美国的MitoSciences公司,加强了免疫分析方面的产品供应;同年也并购了英国的Ascent Scientific 公司,开展了生化试剂的供应。在2012年,Abcam并购了美国的Epitomics 公司,成为一家有的RabMAbs® 供应商。
Abcam 的 目标是给世界上的抗体建立zui大的在线目录,为各地科学家提供*产品,成为各国科学界的重要伙伴。我们为所有产品提供来使客户获得预期的结 果。为提供高质量的抗体来指向尽可能多的靶蛋白,我们尽所能在尽可能多的应用和物种中检测每种抗体。我们相信诚信才是上策,在我们的上会发布有关我们 每一种产品的尽可能多的信息。
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