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使用敲除细胞株进行验证重组RabMAb

重组Anti-MVP抗体[EPR23594-106] - BSA and Azide free (ab273097)

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Western blot - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)
  • Immunocytochemistry/ Immunofluorescence - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)
  • Flow Cytometry (Intracellular) - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)
  • Immunocytochemistry/ Immunofluorescence - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)
  • Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23594-106] to MVP - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

概述

  • 产品名称

    Anti-MVP抗体[EPR23594-106] - BSA and Azide free
    参阅全部 MVP 一抗

  • 描述

    兔单克隆抗体[EPR23594-106] to MVP - BSA and Azide free

  • 宿主

    Rabbit

  • 经测试应用

    适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
    不适用于: IP

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • WB: A549, A431, Caco-2, HeLa treated with 100U/ml IFN gamma for 24 hours, whole, RAW 264.7, NIH/3T3, C6 and PC-12 whole cell lysate. Human colon, lung and kidney tissue lysate. Mouse colon and kidney tissue lysate. IHC-P: Human cerebrum tissue. Human lung cancer and ovary cancer tissue. ICC/IF: A549 and NIH/3T3 cells. Flow Cyt (intra): A549 cells.

  • 常规说明

    ab273097 is the carrier-free version of ab273093.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab273097于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 99 kDa.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.
说明
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 99 kDa.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.
应用说明
Is unsuitable for IP.

靶标

  • 功能

    Required for normal vault structure. Vaults are multi-subunit structures that may act as scaffolds for proteins involved in signal transduction. Vaults may also play a role in nucleo-cytoplasmic transport. Down-regulates INFG-mediated STAT1 signaling and subsequent activation of JAK. Down-regulates SRC activity and signaling through MAP kinases.

  • 组织特异性

    Present in most normal tissues. Higher expression observed in epithelial cells with secretory and excretory functions, as well as in cells chronically exposed to xenobiotics, such as bronchial cells and cells lining the intestine. Overexpressed in many multidrug-resistant cancer cells.

  • 序列相似性

    Contains 9 MVP (vault) repeats.

  • 结构域

    MVP 3 mediates interaction with PTEN.
    MVP 4 mediates interaction with PARP4.

  • 翻译后修饰

    Phosphorylated on Tyr residues after EGF stimulation.
    Dephosphorylated by PTPN11.

  • 细胞定位

    Cytoplasm. Nucleus > nuclear pore complex. 5% found in the nuclear pore complex. Translocates from the nucleus to the cytoplasm upon EGF treatment.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • LRP antibody
    • Lung resistance related protein antibody
    • Lung resistance-related protein antibody
    • Major vault protein antibody
    • Major vault protein, rat, homolog of antibody
    • MVP antibody
    • MVP_HUMAN antibody
    • testicular secretory protein Li 30 antibody
    • VAULT 1 antibody
    • VAULT1 antibody
    see all

图片

  • Western blot - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)
    Western blot - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)
    All lanes : Anti-MVP antibody [EPR23594-106] (ab273093) at 1/1000 dilution

    Lane 1 : Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
    Lane 2 : MVP knockout HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
    Lane 3 : Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate with Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 99 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab273093).

    Lanes 1-3: Merged signal (red and green). Green - ab273093 observed at 110 kDa. Red - loading control ab8245 (Mouse monoclonal [6C5] to GAPDH) observed at 36 kDa.

    ab273093 Anti-MVP antibody [EPR23594-106] was shown to specifically react with MVP in wild-type HeLa cells in Western blot. Significant decrease (8.7 % of intensity compared to the WT band) of signal was observed when MVP knockout cell line ab264817 (knockout cell lysate ab257544) was used.

    ab273093 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4? overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)

    Immunohistochemical analysis of paraffin-embedded human ovary cancer tissue labeling MVP with ab273093 at 1/4000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human ovary cancer (PMID: 23739867). Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab273093).

  • Immunocytochemistry/ Immunofluorescence - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)
    Immunocytochemistry/ Immunofluorescence - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)

    Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized A549 cells labelling MVP with ab273093 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 µg/ml dilution (Green). Confocal image showing cytoplasmic staining in A549 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 µg/ml dilution (Red). The nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 2 µg/ml dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab273093).

  • Flow Cytometry (Intracellular) - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)
    Flow Cytometry (Intracellular) - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)

    Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A549 (Human lung carcinoma epithelial cell) cells labelling MVP with ab273093 at 1/50 dilution (1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor®488, ab150077) at 1/2000 dilution was used as the secondary antibody. 

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab273093). 

     

     

  • Immunocytochemistry/ Immunofluorescence - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)
    Immunocytochemistry/ Immunofluorescence - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)

    Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 cells labelling MVP with ab273093 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 µg/ml dilution (Green). Confocal image showing cytoplasmic staining in NIH/3T3 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 µg/ml dilution (Red). The nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 2 µg/ml dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab273093).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)

    Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling MVP with ab273093 at 1/4000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human lung cancer (PMID: 22117969). Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab273093).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)

    Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling MVP with ab273093 at 1/4000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in endothelium of human cerebrum (PMID: 14636345). Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab273093).

  • Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)
    Anti-MVP antibody [EPR23594-106] - BSA and Azide free (ab273097)

实验方案

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

文献 (0)

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