Anti-Collagen VII antibody [EPR27012-157]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Collagen VII antibody [EPR27012-157] (AB309143)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human skin staining Collagen VII with ab309143 at a 1/100 dilution, ab307969 anti-Aquaporin 3 used at 1/5000 dilution and ab221155 anti-FLG used at a 1/1000 dilution.

Panel A : merged staining of anti-Collagen VII (green; Opal™520), anti-Aquaporin 3 (magenta; Opal™690) and anti-FLG (gray; Opal™570) on human skin.

Panel B : anti-Collagen VII staining basement membrane in human skin.

Panel C : anti-Aquaporin 3 staining squamous epithelium in human skin.

Panel D : anti-FLG staining maturing squamous cells in human skin.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab309143, ab307969 and ab221155 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.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VII antibody [EPR27012-157] (AB309143)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscl tissue labeling Collagen VII with ab309143 at 1/100 (5.32 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on human skeletal muscle.The section was incubated with ab309143 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VII antibody [EPR27012-157] (AB309143)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Collagen VII with ab309143 at 1/100 (5.32 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on human liver.The section was incubated with ab309143 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VII antibody [EPR27012-157] (AB309143)

Immunohistochemical analysis of paraffin-embedded Human skin tissue labeling Collagen VII with ab309143 at 1/100 (5.32 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human skin basement membrane (PMID : 12244305).The section was incubated with ab309143 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VII antibody [EPR27012-157] (AB309143)

Immunohistochemical analysis of paraffin-embedded Rat skin tissue labeling Collagen VII with ab309143 at 1/100 (5.32 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat skin basement membrane.The section was incubated with ab309143 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VII antibody [EPR27012-157] (AB309143)

Immunohistochemical analysis of paraffin-embedded Mouse cornea tissue labeling Collagen VII with ab309143 at 1/100 (5.32 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse cornea basement membrane (PMID : 29920270).The section was incubated with ab309143 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Collagen VII antibody [EPR27012-157] (AB309143)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cornea (fresh) tissue labeling Collagen VII with ab309143 at 1/5000 (0.1064 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Confocal image showing positive staining on rat cornea. The nuclear counterstain was DAPI (Blue). The section was incubated with ab309143 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.<\p> <\p>

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Collagen VII antibody [EPR27012-157] (AB309143)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cornea (fresh) tissue labeling Collagen VII with ab309143 at 1/5000 (0.1064 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Confocal image showing positive staining on mouse cornea. The nuclear counterstain was DAPI (Blue). The section was incubated with ab309143 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.<\p> <\p>

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Collagen VII antibody [EPR27012-157] (AB309143)

Multiplex immunohistochemistry analysis of Formalin/PFA-fixed paraffin-embedded sections of mouse skin tissue staining with ab309143 at 1/100 dilution, ab317813 at 1/500 dilution, and ab315806 at 1/500 dilution. The secondary used was Opal Polymer HRP Ms + Rb. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. Panel A : merged staining of anti-Collagen VII (green; Opal™520), anti-Langerin (gray; Opal™570) and anti-Loricrin (magenta; Opal™690) on mouse skin. Panel B : anti-Collagen VII staining the dermal–epidermal junction in mouse skin. Panel C : anti-Langerin staining Langerhans cells in mouse skin. Panel D : anti-Loricrin staining terminally differentiated epidermal cells in mouse skin. Nuclear DNA was labeled with DAPI (shown in blue). The section was incubated in three rounds of staining : in the order of ab309143, ab317813 and ab315806 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.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Collagen VII antibody [EPR27012-157] (AB309143)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse skeletal muscle (fresh) tissue labeling Collagen VII with ab309143 at 1/5000 (0.106 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Negative control : confocal image showing no staining on mouse skeletal muscle. The nuclear counterstain was DAPI (Blue). The section was incubated with ab309143 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.<\p> <\p>

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Collagen VII antibody [EPR27012-157] (AB309143)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat skeletal muscle (fresh) tissue labeling Collagen VII with ab309143 at 1/5000 (0.1064 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Negative control : confocal image showing no staining on rat skeletal muscle. The nuclear counterstain was DAPI (Blue). The section was incubated with ab309143 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.<\p> <\p>

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VII antibody [EPR27012-157] (AB309143)

Immunohistochemical analysis of paraffin-embedded Mouse skin tissue labeling Collagen VII with ab309143 at 1/100 (5.32 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse skin basement membrane.The section was incubated with ab309143 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Collagen VII antibody [EPR27012-157] (AB309143)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse skin staining Collagen VII with ab309143 at a 1/100 dilution, ab256787 anti-Cdk7 used at 1/100 dilution and ab315806 anti-Loricrin used at a 1/500 dilution.

Panel A : merged staining of anti-Collagen VII (green; Opal™520), anti-Cdk7 (magenta; Opal™690) and anti-Loricrin (gray; Opal™570) on mouse skin.

Panel B : anti-Collagen VII staining basement membrane in mouse skin.

Panel C : anti-Cdk7 staining nucleus in mouse skin.

Panel D : anti-Loricrin staining terminally differentiated epidermal cells in mouse skin.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab309143, ab256787 and ab315806 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.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• WB

Supplier Data

Western blot - Anti-Collagen VII antibody [EPR27012-157] (AB309143)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : HCT116, HEK-293 (PMID : 23743647). The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 11021599). In lane 1, the lysate was stored at -80? prior to Western Blotting. The bands beneath the target band (320 kDa) are likey to be degradation products. In lanes 2-4, the lysates were freshly made and used for Western Blotting immediately to minimize protein degradation. In Western blot, anti- Vinculin antibody (ab129002) loading control staining at 1/10000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-Collagen VII antibody [EPR27012-157] (ab309143) at 1/1000 dilution

Lane 1:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 40 µg

Lane 2:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HCT116 (human colorectal carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 320 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Collagen VII antibody [EPR27012-157] (AB309143)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : liver (PMID : 11673838), kidney (PMID : 1673838). The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 11021599). The bands beneath the target band (320 kDa) are likely to be degraded target fragments. In Western blot, anti- Vinculin antibody (ab129002) loading control staining at 1/10000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-Collagen VII antibody [EPR27012-157] (ab309143) at 1/1000 dilution

Lane 1:

Mouse skin tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 320 kDa

false

Exposure time: 180s