Anti-Rab5 antibody - Early Endosome Marker (ab18211)

Rabbit polyclonal Rab5 antibody. Validated in WB, IP, IHC, ICC/IF and tested in Mouse, Rat, Dog, Human, Chinese hamster. Cited in 61 publication(s). Independently reviewed in 24 review(s).

Overview

  • Product name
    Anti-Rab5 antibody - Early Endosome Marker
    See all Rab5 primary antibodies
  • Description
    Rabbit polyclonal to Rab5 - Early Endosome Marker
  • Host species
    Rabbit
  • Specificity
    The immunogen used for this product shares 43% homology with Rab5C. Cross-reactivity with this protein has not been confirmed experimentally.
  • Tested applications
    Suitable for: IP, IHC-FrFl, IHC-Fr, ICC/IF, WBmore 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
    • Recombinant Human Rab5 protein (ab62956) can be used as a positive control in WB. 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

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.
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

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 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
    All lanes : 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

References

This product has been referenced in:
See all 71 Publications for this product

Customer reviews and Q&As

Filter by Application

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1-10 of 25 Abreviews

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain section - 50um)
Antigen retrieval step
None
Permeabilization
Yes - PBS-T 3 x 5mn
Specification
Brain section - 50um
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jun 13 2019

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse Podocytes)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
75 µg
Specification
Mouse Podocytes
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 30 2018

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

Abcam user community

Verified customer

Submitted 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

Abcam user community

Verified customer

Submitted 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

Abcam user community

Verified customer

Submitted Jul 03 2014

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

Ms. Alina Deliu

Verified customer

Submitted May 21 2013

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

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Dec 04 2012

Application
Western blot
Sample
Human Cell lysate - whole cell (HepaRG)
Loading amount
50 µg
Specification
HepaRG
Gel Running Conditions
Reduced Denaturing (12)
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C

Mrs. Alina Macovei

Verified customer

Submitted May 09 2011

Application
Western blot
Sample
Chinese Hamster Cell lysate - whole cell (Chinese Hamster Ovary cells (CHO))
Loading amount
20 µg
Specification
Chinese Hamster Ovary cells (CHO)
Gel Running Conditions
Reduced Denaturing (4-12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Dec 13 2010

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HepaRG)
Specification
HepaRG
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C

Mrs. Alina Macovei

Verified customer

Submitted Dec 02 2010

1-10 of 25 Abreviews

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