Anti-Rab5 抗体 - Early Endosome Marker (ab18211)

製品の概要

  • 製品名Anti-Rab5 antibody - Early Endosome Marker
    Rab5 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Rab5 - Early Endosome Marker
  • 特異性he immunogen used for this product shares 43% homology with Rab5C. Cross-reactivity with this protein has not been confirmed experimentally.
  • アプリケーション適用あり: IP, IHC-FrFl, IHC-Fr, ICC/IF, WBmore details
  • 種交差性
    交差種: Mouse, Rat, Dog, Human, Chinese Hamster
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human Rab5.

  • ポジティブ・コントロール
    • 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

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
IP Use at an assay dependent concentration.
IHC-FrFl Use at an assay dependent concentration.
IHC-Fr 1/300.
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).

ターゲット情報

Anti-Rab5 antibody - Early Endosome Marker 画像

  • 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.
  • 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.

    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?)
  • 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.

    See 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.

    See Abreview


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 24 kDa


    Exposure time : 150 seconds

Anti-Rab5 antibody - Early Endosome Marker (ab18211) 使用論文

This product has been referenced in:
  • Suo D  et al. Coronin-1 is a neurotrophin endosomal effector that is required for developmental competition for survival. Nat Neurosci 17:36-45 (2014). WB, ICC/IF ; Rat . Read more (PubMed: 24270184) »
  • Simon NC & Barbieri JT Exoenzyme S ADP-ribosylates Rab5 effector sites to uncouple intracellular trafficking. Infect Immun 82:21-8 (2014). Read more (PubMed: 24101692) »

See all 21 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - whole cell (Miapaca2)
Total protein in input 1000 µg
Treatment EGF 50 ng/ml, for 16 h
Immuno-precipitation step Protein A/G
Specification Miapaca2
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Abcam user community

Verified customer

投稿 Jun 16 2016

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Sample Human Cell (ovarian carcinoma (A2780 cell line))
Specification ovarian carcinoma (A2780 cell line)
Permeabilization Yes - 0.1% triton X for 5 min at RT
Fixative Paraformaldehyde
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投稿 Aug 26 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (hepatocytes)
Specification hepatocytes
Treatment 20 ug/ml poly(I:C)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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投稿 Jul 03 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
Sample Human Cell (Hepatoma)
Specification Hepatoma
Permeabilization Yes - 0.2% Triton X-100
Fixative Formaldehyde
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Ms. Alina Deliu

Verified customer

投稿 May 21 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (Rat astrocytes)
Specification Rat astrocytes
Fixative Paraformaldehyde
Permeabilization Yes - 0.3% TritonX in 0.1% PBS
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Dr. Ruma Raha-Chowdhury

Verified customer

投稿 Dec 04 2012

Thank you for confirming this information and for your help and cooperation with this case.

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I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for 2 vials of ab18211.

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

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Thank you for your reply and for kindly providing this further information and order number.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"