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).

Reviews (25)Q&A (9)References (74)

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 corresponding to Human Rab5 aa 150 to the C-terminus (internal sequence) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab18625)

  • 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

  • Western blot - Anti-Rab5 antibody - Early Endosome Marker (ab18211)
    Western blot - Anti-Rab5 antibody - Early Endosome Marker (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 - Anti-Rab5 antibody - Early Endosome Marker (ab18211)
    Immunocytochemistry/ Immunofluorescence - Anti-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 - Anti-Rab5 antibody - Early Endosome Marker (ab18211)
    Western blot - Anti-Rab5 antibody - Early Endosome Marker (ab18211)
    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?

  • Immunohistochemistry (Frozen sections) - Anti-Rab5 antibody - Early Endosome Marker (ab18211)
    Immunohistochemistry (Frozen sections) - Anti-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.

    See Abreview

  • Immunoprecipitation - Anti-Rab5 antibody - Early Endosome Marker (ab18211)
    Immunoprecipitation - Anti-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.
    For Western blotting an HRP conjugated goat monoclonal to rabbit IgG (1/2000) was used.

    See Abreview

  • Western blot - Anti-Rab5 antibody - Early Endosome Marker (ab18211)
    Western blot - Anti-Rab5 antibody - Early Endosome Marker (ab18211)

References

This product has been referenced in:

See all 74 Publications for this product

Publishing research using ab18211? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 34 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Rab5 antibody - Early Endosome Marker

 Good
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Jun 13 2019

Western blot abreview for Anti-Rab5 antibody - Early Endosome Marker

 Below Average
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Apr 30 2018

Immunoprecipitation abreview for Anti-Rab5 antibody - Early Endosome Marker

 Average
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Jun 16 2016

Immunocytochemistry/ Immunofluorescence abreview for Anti-Rab5 antibody - Early Endosome Marker

 Good
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Aug 26 2014

Western blot abreview for Anti-Rab5 antibody - Early Endosome Marker

 Average
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Jul 03 2014

Immunocytochemistry/ Immunofluorescence

 Good
Abreviews
abreview image
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
Read More

Ms. Alina Deliu

Verified customer

Submitted May 21 2013

Immunocytochemistry/ Immunofluorescence abreview for Anti-Rab5 antibody - Early Endosome Marker

 Excellent
Abreviews
abreview image
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
Read More

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Dec 04 2012

Question

Hi Abcam,

The customer has advised;

I would like a credit note for each of the antibodies that I have.

Thank you for your help

Kind regards,

Read More
Answer

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.

Credit note ID: ######

As usual if you have any further questions regarding this credit note, please contact the accounts department by email at creditcontrol@abcam.com. Please refer to the credit ID number in any correspondence with the accounting department.

I would like to wish the customer good luck with their research. The technical team is always at your service, should you require further expert advice.

Read More

Question

Please see below for the response from the customer;

I finally did the staining with the optimised conditions suggested by Abcam, however I see absolutely no staining at all.

I added 0.2% Tween 20 to my wash and dilution buffers and kept the primary antibody overnight and 4 degrees (1:2000 dilution). I haven't attached any pictures as the background is so faint, that I would have to do a 10 sec exposure to see anything.

If however, Abcam wants a picture, I would be happy to send them some.

Hope this problem can be resolved soon!

Thanks for your help

Read More
Answer

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 arrange afree of charge replacementor credit note in compensation for each antibody.

I look forward to hearing from you with details of how you would like to proceed.

Read More

Question

I have purchased 2 vials of ab18211 recently; in the past this antibody has worked well for WB and IHC. However these recent lot GR68012-1have oddly only worked well for WB but not for IHC; a replacement of a new lot GR71410-1 has again worked as expected for both applications; the same samples, reagents and protocols were used in all cases and only the antibody was different

Read More
Answer

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 our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

1-10 of 34 Abreviews or Q&A

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