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使用敲除细胞株进行验证

Anti-Alkaline Phosphatase,组织Non-Specific抗体[2F4] (ab126820)

Submit a review Q&A (3)References (10)
Western blot - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
  • Western blot - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
  • Immunocytochemistry/ Immunofluorescence - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
  • Western blot - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
  • ELISA - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
  • Flow Cytometry - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)

Key features and details

  • Mouse monoclonal [2F4] to Alkaline Phosphatase, Tissue Non-Specific
  • Suitable for: WB, ELISA, IHC-P, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

概述

  • 产品名称

    Anti-Alkaline Phosphatase,组织Non-Specific抗体[2F4]
    参阅全部 Alkaline Phosphatase, Tissue Non-Specific 一抗

  • 描述

    小鼠单克隆抗体[2F4] to Alkaline Phosphatase,组织Non-Specific

  • 宿主

    Mouse

  • 经测试应用

    适用于: WB, ELISA, IHC-P, Flow Cyt, ICC/IFmore details

  • 种属反应性

    与反应: Human

  • 免疫原

    Recombinant fragment corresponding to Human Alkaline Phosphatase, Tissue Non-Specific. expressed in E.coli
    Database link: P05186

  • 阳性对照

    • Recombinant Human ALPL protein (AA: 18-502), HeLa and NTERA-2 cell lysates. This antibody gave a positive result when used in the following formaldehyde/methanol fixed cell lines: HepG2

  • 常规说明

    This product was changed from ascites to supernatant. Lot no’s high than GR307490-5 are from Tissue Culture Supernatant

    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

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab126820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
WB
1/500 - 1/2000. Detects a band of approximately 57.3, 78.9 kDa (predicted molecular weight: 57, 78.9 kDa).
ELISA
1/10000.
IHC-P
1/200 - 1/1000.
Flow Cyt
1/200 - 1/400.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF
1/100.
说明
WB
1/500 - 1/2000. Detects a band of approximately 57.3, 78.9 kDa (predicted molecular weight: 57, 78.9 kDa).
ELISA
1/10000.
IHC-P
1/200 - 1/1000.
Flow Cyt
1/200 - 1/400.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF
1/100.

靶标

  • 功能

    This isozyme may play a role in skeletal mineralization.

  • 疾病相关

    Hypophosphatasia
    Hypophosphatasia childhood type
    Hypophosphatasia infantile type

  • 序列相似性

    Belongs to the alkaline phosphatase family.

  • 翻译后修饰

    N-glycosylated.

  • 细胞定位

    Cell membrane.
  • Information by UniProt

  • 数据库链接

    • 别名

      • AKP2 antibody
      • Alkaline phosphatase liver/bone/kidney antibody
      • Alkaline phosphatase liver/bone/kidney isozyme antibody
      • Alkaline phosphatase tissue nonspecific isozyme antibody
      • Alkaline phosphatase, tissue-nonspecific isozyme antibody
      • Alkaline phosphomonoesterase antibody
      • Alpl antibody
      • AP TNAP antibody
      • AP-TNAP antibody
      • APTNAP antibody
      • BAP antibody
      • FLJ40094 antibody
      • FLJ93059 antibody
      • Glycerophosphatase antibody
      • HOPS antibody
      • Liver/bone/kidney type alkaline phosphatase antibody
      • MGC161443 antibody
      • MGC167935 antibody
      • PHOA antibody
      • PPBT_HUMAN antibody
      • Tissue non specific alkaline phosphatase antibody
      • Tissue nonspecific ALP antibody
      • TNAP antibody
      • TNSALP antibody
      see all

    图片

    • Western blot - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
      Western blot - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
      All lanes : Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820) at 1/500 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : ALPL knockout HAP1 whole cell lysate
      Lane 3 : Saos2 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 57, 78.9 kDa



      Lanes 1 - 3: Merged signal (red and green). Green - ab126820 observed at 80 kDa. Red - loading control, ab181602, observed at 37 kDa.

      ab126820 was shown to specifically react with ALPL in wild-type HAP1 cells as signal was lost in ALPL knockout cells. Wild-type and ALPL knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab126820 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
      Western blot - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
      Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820) at 1/500 dilution + Recombinant Human protein ALPL (AA: 18-502)

      Predicted band size: 57, 78.9 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
      Immunocytochemistry/ Immunofluorescence - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)

      ICC/IF image of ab126820 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab126820 at 1/100 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in methanol fixed (100%, 5min) hepG2 cells used at 1/100 dilution.

    • Western blot - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
      Western blot - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
      All lanes : Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820) at 1/500 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : NTERA-2 cell lysate

      Predicted band size: 57, 78.9 kDa

    • ELISA - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
      ELISA - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
      Example of standard curves obtained using ab126820. Control antigen 100ng (black), antigen at 10ng (purple), antigen at 50ng (blue), antigen at 100ng (red)
    • Flow Cytometry - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
      Flow Cytometry - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
      Flow cytometric analysis of MCF-7 cells using ab126820 at 1/200 (green) and negative control (red).
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody [2F4] (ab126820)
      ab126820, at 1/200, staining Alkaline Phosphatase, Tissue Non-Specific in paraffin embedded Human ovarian cancer tissues by Immunohistochemistry with DAB staining.

    文献 (10)

    发表研究结果有使用 ab126820？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab126820 被引用在 10 文献中.

    • Huang X  et al. Mechanical Sensing Element PDLIM5 Promotes Osteogenesis of Human Fibroblasts by Affecting the Activity of Microfilaments. Biomolecules 11:N/A (2021). PubMed: 34069539
    • Peng Y  et al. Regulation of the integrin αVβ3- actin filaments axis in early osteogenesis of human fibroblasts under cyclic tensile stress. Stem Cell Res Ther 12:523 (2021). PubMed: 34620239
    • Khan AU  et al. Lamin A/C-Dependent Translocation of Megakaryoblastic Leukemia-1 and β-Catenin in Cyclic Strain-Induced Osteogenesis. Cells 10:N/A (2021). PubMed: 34944031
    • Li Z  et al. Graphene oxide-functionalized nanocomposites promote osteogenesis of human mesenchymal stem cells via enhancement of BMP-SMAD1/5 signaling pathway. Biomaterials 277:121082 (2021). PubMed: 34464823
    • Lu H  et al. Modulatory Role of Silver Nanoparticles and Mesenchymal Stem Cell-Derived Exosome-Modified Barrier Membrane on Macrophages and Osteogenesis. Front Chem 9:699802 (2021). PubMed: 34409016
    • Kim H  et al. A novel 3D indirect co-culture system based on a collagen hydrogel scaffold for enhancing the osteogenesis of stem cells. J Mater Chem B 8:9481-9491 (2020). PubMed: 32996551
    • Bertram KM  et al. Identification of HIV transmitting CD11c+ human epidermal dendritic cells. Nat Commun 10:2759 (2019). PubMed: 31227717
    • Casanellas I  et al. Matrix Nanopatterning Regulates Mesenchymal Differentiation through Focal Adhesion Size and Distribution According to Cell Fate. Biomimetics (Basel) 4:N/A (2019). PubMed: 31242712
    • Kundu B  et al. Mechanical Property of Hydrogels and the Presence of Adipose Stem Cells in Tumor Stroma Affect Spheroid Formation in the 3D Osteosarcoma Model. ACS Appl Mater Interfaces 11:14548-14559 (2019). PubMed: 30943004
    • Parlato M  et al. Human ALPI deficiency causes inflammatory bowel disease and highlights a key mechanism of gut homeostasis. EMBO Mol Med 10:N/A (2018). PubMed: 29567797

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    1-3 of 3 Abreviews or Q&A

    Question

    What is the flow cytometry protocol used to validate this antibody?

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    Answer



    Below is the recommended flow cytometry protocol for use with ab126820:



    Flow Cytometry Protocol

    1. Solutions and Reagents

    1.1. 1X PBS

    1.2. Blocking buffer: 0.5% BSA in 1X PBS

    1.3. Filter buffer: 0.1%Triton-X100 and Blocking buffer

    1.4. The in ice cold 4% paraformaldehyde (1% solution - optional for storing samples)

    1.5. Fluorescently-conjugated secondary antibody (various forms)



    2. Protocol

    2.1. Collect 1x106 cells/sample.

    2.2. Wash cells once with blocking buffer.

    2.3. Fix cells with 4% paraformaldehyde and incubate at 4oC for 30 min.

    2.4. Wash cells once with blocking buffer.

    2.5. Add 0.5 ml filter buffer and incubate at 0 oC for 15 min.

    2.6. Wash cells twice with blocking buffer.

    2.7. Incubate cells in blocking buffer for 10 min at room temperature.

    2.8. Add primary antibody at the appropriate dilution (1/200 - 1/400 for ab126820) and incubate for 30 min at room temperature.

    2.9. Wash twice with blocking buffer and incubate with fluorescently-conjugated secondary antibody for 30 min at room temperature.

    2.10. Wash cells twice with blocking buffer.

    2.11. Re-suspend cells in 1X PBS and analyze on flow cytometry. Samples can be kept in 1% paraformaldehyde at 4oC overnight.



    3.1. Advice: Keep the cells in the dark on ice or at 4 oC in a fridge until your scheduled time for analysis. Analyze the cells on the flow cytometer as soon as possible.

    Read More

    Question

    Phone call requesting information on the concentration of the anti-Alkaline Phosphatase antibody (ab126820) and how the negative control was performed for the flow cytometry experiment presented on the datasheet of ab126820.

    Read More
    Answer

    Thank you for your recent enquiries which have been forwarded to me as my colleague is away from the office today.

    I wanted to contact you to let you know we have just received further information regarding the negative control. I can confirm this negative control was a no primary control.

    I hope this will be helpful. If you have any further questions, please do not hesitate to contact us.

    Read More

    Question

    Phone call requesting information on the concentration of the anti-Alkaline Phosphatase antibody (ab126820) and how the negative control was performed for the flow cytometry experiment presented on the datasheet of ab126820.

    Read More
    Answer

    Thank you for contacting us in relation to the Anti-Alkaline Phosphatase (ab126820).

    I am sorry for the delay in my reply. I can now confirm that the concentration of the antibody is 1 mg/mL. This value can therefore be used to calculate the appropriate dilution of your isotype control.

    I hope this information has been of help. If you have any further questions, please do not hesitate to ask.

    Read More

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