遺伝子ノックアウト細胞株 検証済
リコンビナント
RabMAb

Anti-RAB7 抗体 [EPR7589] (ab137029)

製品の概要

  • 製品名
    Anti-RAB7 antibody [EPR7589]
    RAB7 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EPR7589] to RAB7
  • 由来種
    Rabbit
  • アプリケーション
    適用あり: WB, Flow Cyt, IHC-P, ICC/IFmore details
    適用なし: IP
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide corresponding to residues near the C terminal of Human RAB7 (UniProt: P51149)

  • ポジティブ・コントロール
    • A375, A431, U87-MG, HT1080, L929, NIH/3T3, Raw 264.7 and C2C12 cell lysates. Human kidney tissue. Flow Cyt: HAP1-WT cells
  • 特記事項

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab137029 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 1/1000 - 1/10000. Predicted molecular weight: 23 kDa.
Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/500.
  • 追加情報
    Is unsuitable for IP.
  • ターゲット情報

    • 機能
      Key regulator in endo-lysosomal trafficking. Governs early-to-late endosomal maturation, microtubule minus-end as well as plus-end directed endosomal migration and positioning, and endosome-lysosome transport through different protein-protein interaction cascades. Plays a central role, not only in endosomal traffic, but also in many other cellular and physiological events, such as growth-factor-mediated cell signaling, nutrient-transportor mediated nutrient uptake, neurotrophin transport in the axons of neurons and lipid metabolism. Also involved in regulation of some specialized endosomal membrane trafficking, such as maturation of melanosomes, pathogen-induced phagosomes (or vacuoles) and autophagosomes. Plays a role in the maturation and acidification of phagosomes that engulf pathogens, such as S.aureus and M.tuberculosis. Plays a role in the fusion of phagosomes with lysosomes. Plays important roles in microbial pathogen infection and survival, as well as in participating in the life cycle of viruses. Microbial pathogens possess survival strategies governed by RAB7A, sometimes by employing RAB7A function (e.g. Salmonella) and sometimes by excluding RAB7A function (e.g. Mycobacterium). In concert with RAC1, plays a role in regulating the formation of RBs (ruffled borders) in osteoclasts. Controls the endosomal trafficking and neurite outgrowth signaling of NTRK1/TRKA. Regulates the endocytic trafficking of the EGF-EGFR complex by regulating its lysosomal degradation.
    • 組織特異性
      Widely expressed; high expression found in skeletal muscle.
    • 関連疾患
      Defects in RAB7A are the cause of Charcot-Marie-Tooth disease type 2B (CMT2B) [MIM:600882]; also known as hereditary motor and sensory neuropathy II (HMSN2). CMT2B is a form of Charcot-Marie-Tooth disease, the most common inherited disorder of the peripheral nervous system. Charcot-Marie-Tooth disease is classified in two main groups on the basis of electrophysiologic properties and histopathology: primary peripheral demyelinating neuropathy or CMT1, and primary peripheral axonal neuropathy or CMT2. Neuropathies of the CMT2 group are characterized by signs of axonal regeneration in the absence of obvious myelin alterations, normal or slightly reduced nerve conduction velocities, and progressive distal muscle weakness and atrophy. CMT2B is clinically characterized by marked distal muscle weakness and a high frequency of foot ulcers, infections and amputations of the toes. CMT2B inheritance is autosomal dominant.
    • 配列類似性
      Belongs to the small GTPase superfamily. Rab family.
    • 細胞内局在
      Late endosome. Lysosome. Cytoplasmic vesicle > phagosome. Melanosome. Cytoplasmic vesicle > phagosome membrane. Co-localizes with OSBPL1A at the late endosome. Found in the ruffled border (a late endosomal-like compartment in the plasma membrane) of bone-resorbing osteoclasts. Recruited to phagosomes containing S.aureus or Mycobacterium.
    • Information by UniProt
    • 参照データベース
    • 別名
      • CMT2B antibody
      • PRO2706 antibody
      • PSN antibody
      • RAB7, member RAS oncogene family antibody
      • RAB7A antibody
      • RAB7A, member RAS oncogene family antibody
      • RAB7A_HUMAN antibody
      • Ras associated protein RAB7 antibody
      • Ras related protein Rab7 antibody
      • Ras related protein Rab7a antibody
      • Ras-related protein Rab-7a antibody
      see all

    画像

    • Overlay histogram showing HAP1 wildtype (green line) and HAP1-RAB7 knockout cells (red line) stained with ab137029. The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab137029, 0.1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/2000 dilution for 30 min at 22°C.

      A rabbit IgG1 isotype control antibody  (ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-RAB7 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

      Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

      This antibody can also be used in HAP1 cells fixed with 80% methanol (5 min) permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

    • Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling RAB7 with purified ab137029 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

    • Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: RAB7 knockout HAP1 cell lysate (20 µg)
      Lane 3: A431 cell lysate (20 µg)
      Lane 4: Human fetal brain tissue lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab137029 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab137029 was shown to specifically react with RAB7 in wild-type HAP1 cells. No band was observed when RAB7 knockout samples were examined. Wild-type and RAB7 knockout samples were subjected to SDS-PAGE. ab137029 and ab8245 (loading control to GAPDH) were both diluted 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 hour at room temperature before imaging.

    • Flow Cytometry analysis of A431(human epidermoid carcinoma) cells labeling RAB7 with purified ab137029 at 1/20 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

    • ab137029 staining RAB7 in Human HepaRG cell by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.1% Triton X-100 in PBS and blocked with 1% milk for 30 minutes at room temperature. Samples were incubated with primary antibody (1/200 in 1% milk) for 30 minutes. An Alexa Fluor®488-conjugated Donkey anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.

      See Abreview

    • All lanes : Anti-RAB7 antibody [EPR7589] (ab137029) at 1/1000 dilution

      Lane 1 : A375 cell lysate
      Lane 2 : A431 cell lysate
      Lane 3 : U87 MG cell lysate
      Lane 4 : HT 1080 cell lysate
      Lane 5 : L929 cell lysate
      Lane 6 : NIH 3T3 cell lysate
      Lane 7 : Raw 264.7 cell lysate
      Lane 8 : C2C12 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP conjugated Goat anti Rabbit IgG at 1/2000 dilution

      Predicted band size: 23 kDa

    • Immunohistochemical analysis of paraffin embedded Human kidney tissue labelling RAB7 with ab137029 at 1/50 dilution.

    参考文献

    This product has been referenced in:
    • Jimenez-Orgaz A  et al. Control of RAB7 activity and localization through the retromer-TBC1D5 complex enables RAB7-dependent mitophagy. EMBO J 37:235-254 (2018). Human . Read more (PubMed: 29158324) »
    • Takahashi K  et al. Disruption of Small GTPase Rab7 Exacerbates the Severity of Acute Pancreatitis in Experimental Mouse Models. Sci Rep 7:2817 (2017). Read more (PubMed: 28588238) »

    See all 7 Publications for this product

    レビューと Q&A

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    African green monkey Cell (Kidney)
    Permeabilization
    Yes - 0.1% triton x-100
    Specification
    Kidney
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Fixative
    Methanol
    Username

    Abcam user community

    Verified customer

    投稿 Feb 15 2018

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Western blot
    Sample
    African green monkey Cell lysate - whole cell (Kidney)
    Gel Running Conditions
    Reduced Denaturing (4-12% gel)
    Loading amount
    30 µg
    Specification
    Kidney
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    投稿 Dec 29 2017

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Kidney (HEK293 cells))
    Gel Running Conditions
    Reduced Denaturing (4-12% gel)
    Loading amount
    30 µg
    Specification
    Kidney (HEK293 cells)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    投稿 Dec 29 2017

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Brain)
    Gel Running Conditions
    Reduced Denaturing (4-12% gel)
    Loading amount
    30 µg
    Specification
    Brain
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    投稿 Dec 29 2017

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (liver)
    Specification
    liver
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 37°C
    Fixative
    Paraformaldehyde
    Username

    Dr. shuang liang

    Verified customer

    投稿 Aug 01 2017

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (human umbilical vein endothelial cell)
    Gel Running Conditions
    Non-reduced Denaturing (15% gel)
    Loading amount
    20 µg
    Treatment
    siRNA for 48hrs
    Specification
    human umbilical vein endothelial cell
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: ~25°C
    Username

    Dr. 振洋 余

    Verified customer

    投稿 Mar 28 2017

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Human Umbilical Vein Endothelial Cells)
    Permeabilization
    Yes - 0.5%TRITON x-100 for 20mins
    Specification
    Human Umbilical Vein Endothelial Cells
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 37°C
    Fixative
    Paraformaldehyde
    Username

    Dr. 振洋 余

    Verified customer

    投稿 Mar 24 2017

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Spinal Cord)
    Permeabilization
    Yes - 0.2% Triton X-100
    Specification
    Spinal Cord
    Blocking step
    1%BSA +10%FBS as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Methanol
    Username

    Abcam user community

    Verified customer

    投稿 Oct 04 2016

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Brain)
    Gel Running Conditions
    Reduced Denaturing (12)
    Loading amount
    50 µg
    Specification
    Brain
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    投稿 Sep 05 2016

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Tissue lysate - whole (Brain)
    Gel Running Conditions
    Reduced Denaturing (12)
    Loading amount
    50 µg
    Specification
    Brain
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    投稿 Sep 05 2016

    1-10 of 14 Abreviews or Q&A

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

    登録