Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR17077] to Collagen VI - C-terminal
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Collagen VI antibody [EPR17077] - C-terminal
    See all Collagen VI primary antibodies

  • Description

    Rabbit monoclonal [EPR17077] to Collagen VI - C-terminal

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IHC-P
    Mouse
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human fetal heart, skeletal muscle and WI-38 (Human lung) whole cell lysate. Mouse heart, kidney and spleen. Rat heart, kidney and spleen. NIH/3T3 whole cell lysate. IHC-P: Human colon and Mouse cardiac muscle tissue. ICC/IF: NIH/3T3 cells.

  • General notes

     

     

    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.

Properties

Images

  • Western blot - Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720)
    Western blot - Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720)
    All lanes : Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293T whole cell lysate
    Lane 2 : COL6A1 knockout HEK-293T whole cell lysate
    Lane 3 : Human Skeletal Muscle whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 109 kDa



    Lanes 1 - 3: Merged signal (red and green). Green - ab199720 observed at 109 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab199720 was shown to recognize Collagen VI in wild-type Hek 293T cells as signal was lost at the expected MW in COL6A1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and COL6A1 knockout samples were subjected to SDS-PAGE. Ab199720 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720)
    Western blot - Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720)
    All lanes : Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720) at 1/10000 dilution

    Lane 1 : Human fetal heart
    Lane 2 : Human skeletal muscle
    Lane 3 : WI-38 (Human lung) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 109 kDa
    Observed band size: 147 kDa
    why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    Blocking/dilution buffer: 5% NFDM/TBST. 

    The observed MW is consistent with what has been described in the following literature: PMID:16130093 and 21186846

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720)

    Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Collagen VI with ab199720 at 1/1600 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on stromal cells of Human colon tissue is observed. Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720)
    Western blot - Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720)
    All lanes : Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720) at 1/1000 dilution

    Lane 1 : Mouse heart
    Lane 2 : Mouse kidney
    Lane 3 : Mouse spleen
    Lane 4 : Rat heart
    Lane 5 : Rat kidney
    Lane 6 : Rat spleen
    Lane 7 : NIH/3T3 (mouse embryo) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 109 kDa
    Observed band size: 147 kDa why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocking/dilution buffer: 5% NFDM/TBST. 

    The observed MW is consistent with what has been described in the following literature: PMID:16130093 and 21186846

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720)

    Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling Collagen VI with ab199720 at 1/1600 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Extracellular matrix staining on mouse cardiac muscle tissue is observed. Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720)
    Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720)

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