Anti-Rab5抗体- Early Endosome Marker (ab18211)

概述

  • 产品名称
    Anti-Rab5抗体- Early Endosome Marker
    参阅全部 Rab5 一抗
  • 描述
    兔多克隆抗体to Rab5 - Early Endosome Marker
  • 特异性
    he immunogen used for this product shares 43% homology with Rab5C. Cross-reactivity with this protein has not been confirmed experimentally.
  • 经测试应用
    适用于: IP, IHC-FrFl, IHC-Fr, ICC/IF, WBmore details
  • 种属反应性
    与反应: 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.

  • 阳性对照
    • 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

性能

应用

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.

应用 Ab评论 说明
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).

靶标

图片

  • 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

文献

This product has been referenced in:
  • Haghani A  et al. Edible bird's nest modulate intracellular molecular pathways of influenza A virus infected cells. BMC Complement Altern Med 17:22 (2017). Read more (PubMed: 28056926) »
  • Luo Y  et al. Macrophagic CD146 promotes foam cell formation and retention during atherosclerosis. Cell Res 27:352-372 (2017). Read more (PubMed: 28084332) »

See all 44 Publications for this product

客户评价及客户问答

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
Username

Abcam user community

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 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
Username

Ms. Alina Deliu

Verified customer

提交于 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

提交于 Dec 04 2012

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

As requested, I have asked our accounting department to issue you with a credit note. This can then be redeemed against the invoice of a future order...

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Thank you for your message and for providing this further information.

I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments and would be pleased to ...

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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.

To check the status of the order please contact ou...

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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.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

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

I look forward to hearing from you with the results from the next experiment.

Thank you for your time and cooperation. Please do not hesi...

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

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