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Anti-Collagen VI antibody [EPR17072] ab182744 is a rabbit monoclonal antibody that is used in Collagen VI western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone EPR17072 is the most widely used clone for Collagen VI on the market and is cited in >30 publications
- Specificity confirmed with COL6A1 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation

New 20 ul size available


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17072] (AB182744), expandable thumbnail
  • Western blot - Anti-Collagen VI antibody [EPR17072] (AB182744), expandable thumbnail
  • Western blot - Anti-Collagen VI antibody [EPR17072] (AB182744), expandable thumbnail
  • Multiplex immunohistochemistry - Anti-Collagen VI antibody [EPR17072] (AB182744), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Collagen VI antibody [EPR17072] (AB182744), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBICC/IFmIHCIHC-P
Human
Tested
Tested
Tested
Tested
Mouse
Tested
Expected
Expected
Tested
Rat
Expected
Expected
Expected
Tested

Tested
Tested

Species

Mouse

Dilution info

1/2000

Notes

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

Species

Human

Dilution info

1/2000

Notes

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

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/200

Notes

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

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/500 - 1/1000

Notes

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

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/250

Notes

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

Species

Rat

Dilution info

1/250

Notes

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

Species

Human

Dilution info

1/250

Notes

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

Associated Products

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1 product for Alternative Version

Target data

Function

Collagen VI acts as a cell-binding protein.

Alternative names

Collagen alpha-1(VI) chain, COL6A1

Recommended products

Anti-Collagen VI antibody [EPR17072] ab182744 is a rabbit monoclonal antibody that is used in Collagen VI western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone EPR17072 is the most widely used clone for Collagen VI on the market and is cited in >30 publications
- Specificity confirmed with COL6A1 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation

New 20 ul size available

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR17072

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

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

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Collagen VI also known as Collagen 6 is a type of collagen that plays an important role in the extracellular matrix. It consists of three different chains: α1(VI) α2(VI) and α3(VI) with a combined mass of approximately 140 kDa. This collagen type strongly associates with several tissues notably in skeletal muscle skin and the walls of blood vessels. Scientists often study collagen VI with specific collagen antibodies to understand its unique structural properties and its end which binds to other matrix components.

Biological function summary

Collagen VI contributes to structural integrity and cellular scaffolding within tissues. This protein is integral to the formation of microfibrillar networks part of a larger complex that connects the extracellular matrix to cells and other matrix components. Its protective role helps cells resist mechanical stress and the network provided by collagen VI powder stabilizes the tissue architecture. Additionally low endotoxin collagen is often preferable in research to minimize inflammatory responses during experimentation providing more accurate results.

Pathways

Collagen VI participates in extracellular matrix organization and mechanotransduction pathways. It functions alongside other proteins like collagen I and IV. In particular collagen VI interacts with cell surface receptors which influences cellular communication and signaling pathways. This interaction plays a role in transmitting mechanical signals that affect cellular behavior and function proving important for tissue maintenance and repair processes.

Associated diseases and disorders

Mutations or malfunctions involving collagen VI can lead to significant clinical outcomes. It is prominently linked with disorders such as Ullrich congenital muscular dystrophy and Bethlem myopathy both affecting muscle and connective tissue. These conditions often involve disruptions in the assembly or function of the collagen VI network. Additionally related proteins like collagen I and integrin proteins can influence the progression and severity of these disorders illustrating the interconnected nature of these extracellular elements.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

14 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17072] (ab182744), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17072] (ab182744)

    Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Collagen VI with ab182744 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Positive staining around sinusoidal endothelial basement membranes is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-Collagen VI antibody [EPR17072] (ab182744), expandable thumbnail

    Western blot - Anti-Collagen VI antibody [EPR17072] (ab182744)

    Lanes 1-3: Merged signal (red and green). Green - ab182744 observed at 136 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.

    ab182744 Anti-Collagen VI antibody [EPR17072] was shown to specifically react with Collagen VI antibody in wild-type HEK293T cells. Loss of signal was observed when knockout cell line Human COL6A1 knockout HEK-293T cell line ab265060 (knockout cell lysate Human COL6A1 knockout HEK-293T cell lysate ab256879) was used. Wild-type and Collagen VI antibody knockout samples were subjected to SDS-PAGE. ab182744 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-Collagen VI antibody [EPR17072] (ab182744) at 1/1000 dilution

    Lane 1: Wild-type HEK293T cell lysate at 20 µg

    Lane 2: COL6A1 knockout HEK293T cell lysate at 20 µg

    Lane 3: Human skeletal muscle tissue lysate at 20 µg

    Secondary

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

    Predicted band size: 109 kDa

    Observed band size: 136 kDa

    This data was developed using the same antibody clone in a different buffer formulation (ab182744).

    Lanes 1-3: Merged signal (red and green). Green - ab182744 observed at 136 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.

    ab182744 Anti-Collagen VI antibody [EPR17072] was shown to specifically react with Collagen VI antibody in wild-type HEK293T cells. Loss of signal was observed when knockout cell line Human COL6A1 knockout HEK-293T cell line ab265060 (knockout cell lysate Human COL6A1 knockout HEK-293T cell lysate ab256879) was used. Wild-type and Collagen VI antibody knockout samples were subjected to SDS-PAGE. ab182744 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Collagen VI antibody [EPR17072] (ab182744), expandable thumbnail

    Western blot - Anti-Collagen VI antibody [EPR17072] (ab182744)

    Lanes 1 - 3: Merged signal (red and green). Green - ab182744 observed at 109 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.

    ab182744 was shown to recognize Collagen VI in wild-type HEK293 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. ab182744 and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-Collagen VI antibody [EPR17072] (ab182744)

    Lane 1: Wild-type HEK293 whole cell lysate at 20 µg

    Lane 2: COL6A1 knockout HEK293 whole cell lysate at 20 µg

    Lane 3: Human Skeletal Muscle whole cell lysate at 20 µg

    Predicted band size: 105 kDa, 109 kDa

  • Multiplex immunohistochemistry - Anti-Collagen VI antibody [EPR17072] (ab182744), expandable thumbnail

    Multiplex immunohistochemistry - Anti-Collagen VI antibody [EPR17072] (ab182744)

    Multiplex immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue.

    Panel A: Merged staining of Collagen VI (ab182744; green), anti-CD68 (Anti-CD68 antibody [EPR20545] ab213363; red) and anti-Lamin B1 (Anti-Lamin B1 antibody [EPR22165-121] ab229025; magenta).

    Panel B: Anti-Collagen VI (green) stained on extracellular matrix.

    Panel C: Anti-CD68 (red) stained on Kupffer cells.

    Panel D: Anti-Lamin B1 (magenta) stained on nuclear envelope.

    Key protocol steps: The section was incubated in three rounds of staining with ab182744 (1/1000 dilution), Anti-CD68 antibody [EPR20545] ab213363 (1/1000 dilution) and Anti-Lamin B1 antibody [EPR22165-121] ab229025 (1/4000 dilution) for 30 mins at room temperature. Each round was followed by tyramide signal amplification with the appropriate fluorophore.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    DAPI was used as a nuclear counter stain. A ready-to-use anti-Rabbit and Mouse Polymer HRP was used as a secondary.

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-Collagen VI antibody [EPR17072] (ab182744), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Collagen VI antibody [EPR17072] (ab182744)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Collagen VI with ab182744 at 1/200 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm staining on HeLa cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

    The negative controls are as follows:-

    -ve control1 - ab182744 at 1/200 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2 - Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17072] (ab182744), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17072] (ab182744)

    Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling Collagen VI with ab182744 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Positive staining on Human cardiac sarcolemma and interstitium is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17072] (ab182744), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17072] (ab182744)

    Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Collagen VI with ab182744 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Positive staining around basement membranes of Mouse renal tubules is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17072] (ab182744), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17072] (ab182744)

    Immunohistochemical analysis of paraffin-embedded Rat stomach tissue labeling Collagen VI with ab182744 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Positive staining around Rat gastric epithelial basement membranes is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-Collagen VI antibody [EPR17072] (ab182744), expandable thumbnail

    Western blot - Anti-Collagen VI antibody [EPR17072] (ab182744)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Due to posttranslational modifications, observed MW is greater than the predicted MW.

    All lanes: Western blot - Anti-Collagen VI antibody [EPR17072] (ab182744) at 1/20000 dilution

    Lane 1: Human skeletal muscle lysate at 20 µg

    Lane 2: WI-38 (Human fetal lung fibroblast cells) whole cell lysate at 20 µg

    Lane 3: Human placenta lysate at 20 µg

    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

  • Western blot - Anti-Collagen VI antibody [EPR17072] (ab182744), expandable thumbnail

    Western blot - Anti-Collagen VI antibody [EPR17072] (ab182744)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Due to posttranslational modifications, observed MW is greater than the predicted MW.

    All lanes: Western blot - Anti-Collagen VI antibody [EPR17072] (ab182744) at 1/2000 dilution

    Lane 1: Human fetal brain lysate at 10 µg

    Lane 2: Human fetal heart lysate at 10 µg

    Lane 3: Human fetal kidney lysate at 10 µg

    Lane 4: Human fetal spleen lysate at 10 µg

    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

  • Western blot - Anti-Collagen VI antibody [EPR17072] (ab182744), expandable thumbnail

    Western blot - Anti-Collagen VI antibody [EPR17072] (ab182744)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Due to posttranslational modifications, observed MW is greater than the predicted MW.

    All lanes: Western blot - Anti-Collagen VI antibody [EPR17072] (ab182744) at 1/2000 dilution

    Lane 1: Mouse heart lysate at 10 µg

    Lane 2: Mouse kidney lysate at 10 µg

    Lane 3: Mouse spleen lysate at 10 µg

    Lane 4: Rat kidney lysate at 10 µg

    Lane 5: Rat spleen lysate at 10 µg

    Lane 6: NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 µg

    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

  • Multiplex immunohistochemistry - Anti-Collagen VI antibody [EPR17072] (ab182744), expandable thumbnail

    Multiplex immunohistochemistry - Anti-Collagen VI antibody [EPR17072] (ab182744)

    Fluorescence multiplex immunohistochemical analysis of human prostate gland tissue (formalin/PFA-fixed paraffin-embedded section). Panel A: merged staining of anti-Collagen VI (Anti-Collagen VI antibody [EPR17072] - BSA and Azide free ab271938, magenta; Opal™690), anti-Cytokeratin 5 (Anti-Cytokeratin 5 antibody [SP27] - BSA and Azide free ab236216, green; Opal™520) and anti-Prostate Specific Antigen (Anti-Prostate Specific Antigen antibody [EP1588Y] ab76113, red; Opal™570) on human prostate gland tissue. Panel B: anti-Prostate Specific Antigen stained on luminal cells. Panel C: anti-Cytokeratin 5 stained on basal cells. Panel D: anti-Collagen VI stained on stroma. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of Anti-Collagen VI antibody [EPR17072] - BSA and Azide free ab271938 (1/500), Anti-Cytokeratin 5 antibody [SP27] - BSA and Azide free ab236216 (1/400), and Anti-Prostate Specific Antigen antibody [EP1588Y] ab76113 (1/2000) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) was used for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

  • Multiplex immunohistochemistry - Anti-Collagen VI antibody [EPR17072] (ab182744), expandable thumbnail

    Multiplex immunohistochemistry - Anti-Collagen VI antibody [EPR17072] (ab182744)

    This data was developed using the same antibody clone in a different buffer formulation (Anti-Collagen VI antibody [EPR17072] - BSA and Azide free ab271938).

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast labelling Cytokeratin 18 with Anti-Cytokeratin 18 antibody [SP69] ab93741 at 1/200 dilution (1.02 µg/mL) (B), Collagen VI with Anti-Collagen VI antibody [EPR17072] - BSA and Azide free ab271938 at 1/500 dilution (2.084 μg/ml) (C) and Cytokeratin 14 with Anti-Cytokeratin 14 antibody [SP53] - BSA and Azide free ab236439 at 1/2000 dilution ( 0.519 μg/ml) (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

    Panel A: merged staining of anti-Cytokeratin 14 (magenta; Opal™690), anti-Cytokeratin 18 (green; Opal™520) and anti-Collagen VI (red; Opal™570) on human breast.
    Panel B: anti-Cytokeratin 18 stained on luminal epithelial cells.
    Panel C: anti-Collagen VI stained on stroma.
    Panel D: anti-Cytokeratin 14 stained on myoepithelial cells.

    The section was incubated in three rounds of staining: in the order of Anti-Cytokeratin 14 antibody [SP53] - BSA and Azide free ab236439 for 30 mins, Anti-Cytokeratin 18 antibody [SP69] ab93741 for 10 mins, and Anti-Collagen VI antibody [EPR17072] - BSA and Azide free ab271938 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

  • Multiplex immunohistochemistry - Anti-Collagen VI antibody [EPR17072] (ab182744), expandable thumbnail

    Multiplex immunohistochemistry - Anti-Collagen VI antibody [EPR17072] (ab182744)

    Fluorescence multiplex immunohistochemical analysis of human adrenal gland (formalin-fixed paraffin-embedded section). Panel A shows merged staining of anti-PCSK2 stained on adrenal medulla (Anti-PCSK2 antibody [EPR23578-19] ab274418; gray; Opal™690) at 1:2000 (0.263 μg/ml) [Panel B], anti-CYP11A1 stained on adrenal cortex (Anti-CYP11A1 antibody [EPR24868-86] ab272494; green; Opal™520) at 1:10000 (0.053 μg/ml) [Panel C], and anti-Collagen VI stained on extracellular matrix (ab182744; red; Opal™570) at 1:1000 (1.518 μg/ml) [Panel D] on human adrenal gland. DAPI was used as a nuclear counter stain. Followed by Opal Polymer HRP Ms + Rb secondary antibody. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. The section was incubated in three rounds of staining: in the order of Anti-PCSK2 antibody [EPR23578-19] ab274418, Anti-CYP11A1 antibody [EPR24868-86] ab272494, and ab182744 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used. 

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