Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online

Anti-Rab5 抗体 (ab13253)

Reviews (3)Q&A (3)References (32)
Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody (ab13253)
  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody (ab13253)
  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody (ab13253)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody (ab13253)

Key features and details

  • Rabbit polyclonal to Rab5
  • Suitable for: IHC-P, ICC/IF
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

リコンビナント抗体で、ロット間での高い再現性を実現

Product image
Anti-Rab5 antibody [EPR21801] - Early Endosome Marker (ab218624)
  • 異なるロット間での安定した再現性
  • 容易なスケールアップ
  • 評価試験による特異性の確認済み
  • 倫理基準に準拠 - アニマル・フリーの生産

製品の概要

  • 製品名

    Anti-Rab5 antibody
    Rab5 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to Rab5

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: IHC-P, ICC/IFmore details

  • 種交差性

    交差種: Mouse, Rat, Human
    交差が予測される動物種: Dog

  • 免疫原

    Synthetic peptide corresponding to Human Rab5 aa 150-250 conjugated to keyhole limpet haemocyanin.
    Database link: P20339

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • ポジティブ･コントロール

    • ICC/IF: Rat hippocampal neurons (embryonic day 18), HEK cells, HeLa cells. IHC-P: Mouse back skin tissue section.

  • 特記事項

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab13253の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
IHC-P
1/100.

Use Bouin's fixative to fix tissue.
ICC/IF
1/80.
特記事項
IHC-P
1/100.

Use Bouin's fixative to fix tissue.
ICC/IF
1/80.

ターゲット情報

画像

  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody (ab13253)
    Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody (ab13253)This image is courtesy of Randal Moldrich, CNRS UMR7637, ESPCI, France
    Confocal microscopy of fixed primary cultures of rat hippocampal neurons (embryonic day 18) showing immunocytochemical labelling of polyclonal rabbit anti-Rab5 (Abcam ab13253, 1/200; Alexa Fluor 488; 1/200; green) and monoclonal mouse anti-tubulin-II (Alexa Fluor 568 1/200; red). Magnification 63x; insert 180x.
  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody (ab13253)
    Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody (ab13253)This image is courtesy of Randal Moldrich, CNRS UMR7637, ESPCI, France

    ab13253 Rabbit polyclonal to Rab5 was used in in fixed HEK cells that had been transfected with Rab5-GFP Fluorescence microscopy of fixed Rab5-transfected HEK cells showing (left to right) immunocytochemical labelling of rabbit polyclonal Rab5 (ab13253, 1/500; Cy3; red), and Rab5-GFP (green) with their overlay (far right). Scale bar = 7µm. Showing good co-localisation.

  • Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody (ab13253)
    Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody (ab13253)

    Immunofluorescent analysis of 2% paraformaldehyde fixed Human cervical cancer cell line (HeLa) for 20min at RT, labelling Rab5 with ab13253 at 1/80 dilution for 12 hours at 4°C, followed by secondary antibody Goat Anti-Rabbit (FITC) (green) at 1/200 dilution for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Image showing cytoplasmic staining on HeLa cells. (A) DAPI (blue) nuclear stain. (B) Anti-Rab5 Antibody (green). (C) Composite image.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody (ab13253)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab5 antibody (ab13253)

    ab13253 at 1/100 dilution staining Rab5 in mouse back skin tissue section by Immunohistochemistry (Bouin's fixative fixed paraffin-embedded tissue section). Antigen retrieval was done by microwave in citrate buffer. A Fluorophore-conjugated goat anti-rabbit secondary was used at 1/50 dilution.

参考文献 (32)

ab13253 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab13253 は 32 報の論文で使用されています。

  • Bagalkot TR  et al. Dopamine Transporter Localization in Medial Forebrain Bundle Axons Indicates Its Long-Range Transport Primarily by Membrane Diffusion with a Limited Contribution of Vesicular Traffic on Retromer-Positive Compartments. J Neurosci 41:234-250 (2021). PubMed: 33234607
  • Kusama K  et al. Day 7 Embryos Change the Proteomics and Exosomal Micro-RNAs Content of Bovine Uterine Fluid: Involvement of Innate Immune Functions. Front Genet 12:676791 (2021). PubMed: 34262596
  • Han J  et al. Endocytic BDNF secretion regulated by Vamp3 in astrocytes. Sci Rep 11:21203 (2021). PubMed: 34707216
  • Puangmalai N  et al. Internalization mechanisms of brain-derived tau oligomers from patients with Alzheimer's disease, progressive supranuclear palsy and dementia with Lewy bodies. Cell Death Dis 11:314 (2020). PubMed: 32366836
  • Carim SC  et al. IPIP27 Coordinates PtdIns(4,5)P2 Homeostasis for Successful Cytokinesis. Curr Biol 29:775-789.e7 (2019). PubMed: 30799246
View all Publications for this product

レビューと Q&A

Show All レビュー Q&A
レビューを送る 質問を送る

1-6 of 6 Abreviews or Q&A

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

 Average
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - other (HeLa)
Loading amount
100000 cells
Specification
HeLa
Gel Running Conditions
Reduced Denaturing (7.5%)
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Mar 31 2009

Immunocytochemistry abreview for Anti-Rab5 antibody - Early Endosome Marker

 Average
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Rat Cell (Hippocampal neuron culture)
Specification
Hippocampal neuron culture
Fixative
Paraformaldehyde
Blocking step
BSA as blocking agent for 40 minute(s) · Concentration: 5%
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Randal Moldrich

Verified customer

投稿 Dec 02 2005

Immunocytochemistry abreview for Anti-Rab5 antibody - Early Endosome Marker

 Good
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Human Cell (human Rab5-transfected HEK cells)
Specification
human Rab5-transfected HEK cells
Fixative
Methanol
Blocking step
BSA as blocking agent for 40 minute(s) · Concentration: 5%
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Randal Moldrich

Verified customer

投稿 Dec 02 2005

Question

Product code: 13253
Inquiry: Dear sir/ madam, I am looking for a primary antibodies that reacts with mouse Rab5 for the specific use of immunofluorescence analysis. I was wondering if you have details on whether product code ab13253 have been used in immunofluorescence studies on mouse cells? Thanks.

Read More
Answer

Thank you for contacting us.

The antibody ab13253 is guaranteed for ICC/IF in mouse cells; this means this product comes with fully money back guarantee if used as stated on the datasheet.

We unfortunately do not have any mouse specific image however we do have tested this with mouse cells so please buy the ab without any hesitation.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Question

I am seeing nuclear staining with this antibody, when it is listed as being localized at the cytoplasmic surface of the plasma membrane. Have you seen nuclear staining with this antibody before?

Read More
Answer

Thank you for your patience, and I apologize for the delay in answering your question. We have not personally tested this antibody for IHC/IF but it has been cited in a publication: http://www.jbc.org/cgi/content/full/276/21/18540 As one would expect with Rab5, the staining in the publication is localized to the membrane region of the tissue. The protocol used is well detailed in the publication, including their antigen unmasking technique. It is possible that you will get better staining by optimizing your procedure a bit. The nuclear staining may just be non-specific. I hope this information helps. Please do not hesitate to contact us if you need anything further.

Read More

Question

BATCH NUMBER 74988 DESCRIPTION OF THE PROBLEM High background. No signal SAMPLE Cos 7 cells, whole cell, light mitochondrial pellet or subcellular fractions. PRIMARY ANTIBODY rabbit anti Rab5 abcam, 1:2000 SECONDARY ANTIBODY goat anti rabbit, peroxidase coupled, Jackson DETECTION METHOD ECL, amersham POSITIVE AND NEGATIVE CONTROLS USED positive control: I saw a band for Rab 5 once with whole cell lysate. All other attempts with this antibody resulted in no signal after shorter exposures or black blots after longer exposures. negative control: only secondary antibody gives no background other positive controls: a rabbit anti calreticulin antibody I use works fine. The same secondary antibody is used by many other people in the lab and works fine. All the other antibodies I use (mouse anti Tom22, anti cyt c oxidase, NaK ATPase, EEA1, LAMP-1; rat anti HA) work great without background. ANTIBODY STORAGE CONDITIONS Arrived at RT. Aliquoted on day of arrival. 10 micro liter aliquots stored at -20. Used aliquots stored at 4 degrees. SAMPLE PREPARATION Differential Centrifugation - 3 60mm Petri dishes of COS 7 cells - rinse cells once with cold PBS, scrape cells into 2ml cold PBS per dish - spin cells down 1min on setting 4 - resuspend pellet in 500microl cold HM, transfer to Eppendorf vial, spin 1min at 5000g - take off supernatant, resuspend pellet in 100microl cold HM + complete - transfer to cold Dounce homogenizer, homogenize with 25 strokes using pestle B - add 300microl cold HM + complete, transfer to Eppendorf tube, spin 10min at 1000g, 4?C - save supernatant, resuspend pellet in 100microl cold HM + complete - transfer to cold Dounce homogenizer, homogenize with 25 strokes using pestle B - add 300microl cold HM + complete, transfer to Eppendorf tube, spin 10min at 1000g, 4?C - take off supernatant, combine supernatants, spin 10min at 1000g, 4?C - take off supernatant, spin 10min at 1000g, 4?C - take off supernatant, spin 10min at 17,000g, 4?C -> resuspend pellet in 1.5ml 30% OptiPrep, save 250microl for Western - layer under OptiPrep gradient - centrifuge 3 hours at 52,000gav (20 500rpm for TH-641 rotor) in OptiPrep gradient. - collect 13 drop fractions - mix each fractions with 4x SDS sample buffer (with BME) - load 15 microl of each fraction and of input for Western blot AMOUNT OF PROTEIN LOADED Not determined. ELECTROPHORESIS/GEL CONDITIONS 9% SDS Tris gels. BME as reducing agent. TRANSFER AND BLOCKING CONDITIONS NaCO3/NaHCO3 buffer with 10% methanol. PVDF membrane. Block: 1% milkpowder in TBS + 0.5% Tween. 1h at RT or over night at 4 degrees. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 6 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? - higher and lower secondary concentration (1:2000, 1:5000, 1:10,000) - more washes - longer and shorter exposure times (20s to 1 hour) ADDITIONAL NOTES I did have similar problems with the rabbit anti-calreticulin antibody I use. But this problem was solved by altering the secondary concentration.

Read More
Answer

Thank you for your enquiry and for taking your time to fill in the on-line questionnaire. We are very sorry to hear that you are having problem with this antibody. According to our order record and the database, we have sold several vials of this batch number without any problem. Therefore, at this stage, we would suggest that there is either a problem with the vial you received, or modifications to your protocol are needed to obtain a positive result. We would strongly recommend determining the protein concentration of the samples and loading at least 25 or 30 ug per lane. Otherwise you can’t be sure that you have loaded enough material onto the gel. The recommended dilution for Western blot is 1/2000 if sufficient amount of protein is loaded. Since you have not determined the concentration, you may need to increase the concentration of the primary antibody (1/500 and 1/1000) to see if the signal is getting stronger. You have not mentioned how long you incubated the membrane with the primary antibody. You can also try to apply overnight incubation at 4 oC or 2 hrs at room temperature. It may also be worth running positive control along with the samples. This antibody has been tested for Western blot application using mouse or rat whole brain tissue extract. Try to block the nonspecific bindings with 5% BSA or 5% nonfat dry milk instead of 1% milk. We hope this will help. Should you still have any problem with this product, then please do not hesitate to contact us again.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com