製品の概要

  • 製品名
  • 製品の詳細
    Mouse monoclonal to RAB27A
  • 由来種
    Mouse
  • アプリケーション
    適用あり: WB, IHC-P, IP, IHC-FoFr, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Human
    交差が予測される動物種: Rabbit
  • 免疫原

    Recombinant fragment: YCENPDIVLC GNKSDLEDQR VVKEEEAIAL AEKYGIPYFE TSAANGTNIS QAIEMLLDLI MKRMERCVDK SWIPEGVVRS NGHASTDQLS EEKEKGACGC , corresponding to amino acids 122-222 of Human RAB27A

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab55667 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
WB Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 25 kDa.
IHC-P Use a concentration of 3 µg/ml.
IP Use at an assay dependent concentration. See Abreview.
IHC-FoFr Use at an assay dependent concentration. PubMed: 21832089
ICC/IF 1/200.

ターゲット情報

  • 機能
    Plays a role in cytotoxic granule exocytosis in lymphocytes. Required for both granule maturation and granule docking and priming at the immunologic synapse.
  • 組織特異性
    Found in all the examined tissues except in brain. Low expression was found in thymus, kidney, muscle and placenta. Detected in melanocytes, and in most tumor cell lines examined. Expressed in cytotoxic T-lymphocytes (CTL) and mast cells.
  • 関連疾患
    Defects in RAB27A are a cause of Griscelli syndrome type 2 (GS2) [MIM:607624]. Griscelli syndrome is a rare autosomal recessive disorder that results in pigmentary dilution of the skin and hair, the presence of large clumps of pigment in hair shafts, and an accumulation of melanosomes in melanocytes. GS2 patients also develop an uncontrolled T-lymphocyte and macrophage activation syndrome, known as hemophagocytic syndrome, leading to death in the absence of bone marrow transplantation. Neurological impairment is present in some patients, likely as a result of hemophagocytic syndrome.
  • 配列類似性
    Belongs to the small GTPase superfamily. Rab family.
  • 細胞内局在
    Membrane. Melanosome. Late endosome. Lysosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. Localizes to endosomal exocytic vesicles.
  • Information by UniProt
  • 参照データベース
  • 別名
    • GS2 antibody
    • GTP-binding protein Ram antibody
    • HsT18676 antibody
    • MGC117246 antibody
    • Mutant Ras related protein Rab-27A antibody
    • Rab-27 antibody
    • RAB-27A antibody
    • RAB27 antibody
    • RAB27A antibody
    • RAB27A member RAS oncogene family antibody
    • RAM antibody
    • Ras-related protein Rab-27A antibody
    • Ras-related protein Rab27A antibody
    • RB27A_HUMAN antibody
    see all

画像

  • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: RAB27A knockout HAP1 whole cell lysate (20 µg)
    Lane 3: Jurkat whole cell lysate (20 µg)
    Lane 4: MCF7 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab55667 observed at 27 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab55667 was shown to specifically react with RAB27A when RAB27A knockout samples were used. Wild-type and RAB27A knockout samples were subjected to SDS-PAGE. Ab55667 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 2.5 ug/ml and 1/10000 dilution respectively. Blots were developed with IRDye® 800CW Goat anti-Mouse IgG (H + L) and IRDye® 680 Goat anti-Rabbit IgG (H + L) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Anti-RAB27A antibody (ab55667) + HL-60 cell lysate

    Predicted band size: 25 kDa

  • Immunocytochemistry/ Immunofluorescence  analysis of SK-MEL-28 cells labeling RAB27A with ab55667 at 1/200 dilution. The cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton-X in PBS. A polyclonal goat anti-mouse IgG Alexa Fluor® 488 secondary antibody was used at 1/1000 dilution.

    See Abreview

  • All lanes : Anti-RAB27A antibody (ab55667)

    Lane 1 : RAB27A over-expressed 293 cell line, cotransfected with RAB27A Validated Chimera RNAi
    Lane 2 : Non-transfected control

    Predicted band size: 25 kDa



    GAPDH (36.1 kDa) used as specificity and loading control.

  • All lanes : Anti-RAB27A antibody (ab55667)

    Lane 1 : RAB27A transfected lysate(24.9 KDa)
    Lane 2 : Non-transfected lysate

    Predicted band size: 25 kDa

  • RAB27A antibody (ab55667) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human lymphoma. Heat induced antigen retrieval with Citrate buffer pH 6 has been performed prior to immunostaining.
  • ab55667 staining cow Retinal Pigment Epithelium (RPE) cells by ICC/IF.  Cells were PFA fixed and permeabilized in 0.5% Triton X-100 prior to blocking in 10% serum for 20 minutes at 25°C.  The primary antibody was diluted 1/200 and incubated with the sample for 12 hours at 4°C.  An Alexa Fluor® 546 conjugated goat anti-mouse antibody was used as the secondary.  Rab27a, which localizes to the apical site of the RPE cells, is labeled with red fluorescence. Nuclei were counterstained with DAPI (blue).

    See Abreview

  • All lanes : Anti-RAB27A antibody (ab55667) at 1/200 dilution

    Lane 1 : whole cell lysate of HEK 293 transfected with empty pCDNA3 vector
    Lane 2 : whole cell lysate of HEK 293 cells expressing Rab27a-GFP

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP conjugated goat anti-mouse

    Developed using the ECL technique.

    Predicted band size: 25 kDa
    Observed band size: 52 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 minutes


    Rab27a (25kDa)+GFP(27kDa)=52kDa

    The blot was blocked with 5% milk for 30 minutes at 25°C and incubated with the primary antibody for 12 hours at 4°C.

    See Abreview

  • ab55667 (undiluted) was incubated with whole cell lysate of HEK 293 cells transfected with Rab27a-GFP and a Protein G matrix for 12 hours at 4°C to achieve immunoprecipitation.  20µg of lysate were present in the input.

    ab55667 (diluted 1/200) was also used in the western blotting step.

    Lane 1. lysate of HEK 293 cells expressing Rab27a-GFP
    Lane 2. IP with ab55667
    Lane 3. Not bound fraction

    See Abreview

参考文献

This product has been referenced in:
  • Chen L  et al. HCC-derived exosomes elicit HCC progression and recurrence by epithelial-mesenchymal transition through MAPK/ERK signalling pathway. Cell Death Dis 9:513 (2018). Read more (PubMed: 29725020) »
  • Jiang C  et al. GGPP-Mediated Protein Geranylgeranylation in Oocyte Is Essential for the Establishment of Oocyte-Granulosa Cell Communication and Primary-Secondary Follicle Transition in Mouse Ovary. PLoS Genet 13:e1006535 (2017). Read more (PubMed: 28072828) »
See all 16 Publications for this product

レビューと Q&A

1-10 of 12 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse hepatocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
mouse hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

投稿 Jul 09 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Human Dermal Microvascular Endothelial Cells)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
50 µg
Specification
Human Dermal Microvascular Endothelial Cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

投稿 Feb 23 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rabbit Tissue sections (intestinum)
Specification
intestinum
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated
Permeabilization
No
Blocking step
Blocking Solution Zytomed as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

投稿 Mar 01 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (kidney)
Specification
kidney
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer
Permeabilization
No
Blocking step
(agent) for 30 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

投稿 Dec 20 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (SK-MEL-28)
Specification
SK-MEL-28
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton-X in PBS

Abcam user community

Verified customer

投稿 Oct 18 2012

Answer

Selon nos informations vous avez connu quelques difficultés avec ab55667 et avez contacté notre service scientifique (en février).

Nous vous avons donné quelques suggestions pour améliorer le protocole. Je vous écris maintenant pour savoir si ces suggestions ont permis de résoudre le problème ou si nous pouvions vous aider autrement? J'aimerais bien aussi vous rappeler que nous avons une garantie sur nos produits, et si nous arrivons donc pas à résoudre le problème, nous allons vous le remplacer.

Je vous souhaite bonne chance dans vos recherches. N’hésitez pas à me recontacter pour plus d’assistance.

Read More

Answer

Merci d'avoir pris le temps de remplir notre questionnaire et de nous avoir contactés. Je suis désolée d'apprendre que vous avez des difficultés à obtenir de bons résultats avec ab55667.

Merci d'avoir patienté pendant que je discutais avec le laboratoire. Notre politique est d'offrir tout d'abord la meilleure assistance technique. Si nous concluons que le produit que vous avez reçu ne fonctionne pas comme mentionné sur sa fiche technique et que ce produit a été commandé sous 180 jours, nous nous ferons un plaisir de vous offrir un remplacement gratuit ou un avoir en compensation.

J'aimerais donc vous offrir quelques suggestions afin d'optimiser vos résultats. J'apprécierais si vous pouviez confirmer quelques détails de la procédure.

1.)Effectivement, je pense qu'il est possible que vous voyez que du bruit de fond. Les anticorps ont tendance à précipiter sur le noyau. Donc à trop forte concentration, il se peux que ce donne un bruit de fond nucléaire. Je peux donc vous conseiller de diluer l'anticorps à 1/200 et 1/400.

2.) Pour obtenir un marquage positive, je vous conseille de utiliser du démasquage à la chaleur avec un tampon Citrate. Effectivement, le laboratoire utilise cet méthode pour détecter Rab27a avec cet anticorps.

J'espère que ces informations vous aideront. Veuillez s'il vous plaît me tenir au courant de comment vous procédez. Veuillez ne pas hésiter non plus à nous recontacter si vous avez des questions.

Read More

Question
Answer

Merci pour votre appel. Je suis désolée d'apprendre que ab55667 ne vous donne pas de bons résultats en immunohistochimie.

Veuillez trouver ci-joint un questionnaire, que nous avons discuté, qui nous permet de regrouper le plus d'informations possible sur le protocole que vous avez utilisé avec cet anticorps. Veuillez s'il vous plaît nous envoyer aussi une image avec les résultats obtenus.
Ceci-ci nous permettra de vous fournir la meilleure assistance technique.

Si nous concluons que notre produit est fautif et ne fonctionne pas comme explicité par notre fiche technique, nous serons ravis de mettre en place l'envoi d'un tube de remplacement gratuit ou d'un remboursement (sous réserve d'une commande ultérieure de 180 jours).

Je vous remercie par avance de nous envoyer votre protocole.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HEK 293 cells transfected with hRab27a-GFP)
Total protein in input
20 µg
Specification
HEK 293 cells transfected with hRab27a-GFP
Immuno-precipitation step
Protein G

Herr Dr. Vladimir Milenkovic

Verified customer

投稿 Feb 29 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK-293 cells transfected with human Rab27a-GFP)
Loading amount
20 µg
Specification
HEK-293 cells transfected with human Rab27a-GFP
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Herr Dr. Vladimir Milenkovic

Verified customer

投稿 Feb 29 2008

1-10 of 12 Abreviews or Q&A

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

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