Validated using a knockout cell lineRecombinantRabMAb

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

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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

    Suitable for: IHC-P, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human Collagen VI aa 800 to the C-terminus. The exact sequence is proprietary.
    Database link: P12109

  • 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab199720 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P 1/800. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 147 kDa (predicted molecular weight: 109 kDa).

Target

  • Function

    Collagen VI acts as a cell-binding protein.

  • Involvement in disease

    Defects in COL6A1 are a cause of Bethlem myopathy (BM) [MIM:158810]. BM is a rare autosomal dominant proximal myopathy characterized by early childhood onset (complete penetrance by the age of 5) and joint contractures most frequently affecting the elbows and ankles.
    Defects in COL6A1 are a cause of Ullrich congenital muscular dystrophy (UCMD) [MIM:254090]; also known as Ullrich scleroatonic muscular dystrophy. UCMD is an autosomal recessive congenital myopathy characterized by muscle weakness and multiple joint contractures, generally noted at birth or early infancy. The clinical course is more severe than in Bethlem myopathy.

  • Sequence similarities

    Belongs to the type VI collagen family.
    Contains 3 VWFA domains.

  • Post-translational
    modifications

    Prolines at the third position of the tripeptide repeating unit (G-X-Y) are hydroxylated in some or all of the chains.

  • Cellular localization

    Secreted > extracellular space > extracellular matrix.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • Alpha 1 (VI) chain (61 AA) antibody
    • CO6A1_HUMAN antibody
    • COL6A1 antibody
    • COL6A2 antibody
    • COL6A3 antibody
    • collagen 6 antibody
    • Collagen alpha 2(VI) chain antibody
    • Collagen alpha 3(VI) chain antibody
    • Collagen alpha-1(VI) chain antibody
    • collagen six antibody
    • Collagen type VI alpha 1 antibody
    • Collagen type VI alpha 2 antibody
    • Collagen type VI alpha 3 antibody
    • Collagen VI alpha 1 polypeptide antibody
    • Collagen VI alpha 2 polypeptide antibody
    • Collagen VI alpha 3 polypeptide antibody
    • CollagenVI antibody
    • Human mRNA for collagen VI alpha 2 C terminal globular domain antibody
    • OPLL antibody
    • PP3610 antibody
    see all

Images

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

References (2)

Publishing research using ab199720? Please let us know so that we can cite the reference in this datasheet.

ab199720 has been referenced in 2 publications.

  • Wang D  et al. Insulin-like growth factor-1 engaged in the mandibular condylar cartilage degeneration induced by experimental unilateral anterior crossbite. Arch Oral Biol 98:17-25 (2019). PubMed: 30419485
  • Endicott J  et al. Authentication of collagen VI antibodies. BMC Res Notes 10:358 (2017). PubMed: 28755659

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1-2 of 2 Abreviews or Q&A

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

 Poor
Abreviews
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Sheep Tissue sections (femoral artery / femoral vein)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer pH 6.0
Permeabilization
Yes - Triton X
Specification
femoral artery / femoral vein
Blocking step
Serum (+ 3% BSA) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1.5% · Temperature: RT°C
Fixative
Paraformaldehyde
Read More

Dr. Jonas Keiler

Verified customer

Submitted Jun 08 2018

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

 Poor
Abreviews
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (femoral artery / femoral vein)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer pH 6.0
Permeabilization
Yes - Triton X
Specification
femoral artery / femoral vein
Blocking step
Serum (+ 3% BSA) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1.5% · Temperature: RT°C
Fixative
Paraformaldehyde
Read More

Dr. Jonas Keiler

Verified customer

Submitted Jun 08 2018

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