Anti-Collagen VI antibody [EPR17072] - BSA and Azide free

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Collagen VI antibody [EPR17072] - BSA and Azide free (AB271938)

Fluorescence multiplex immunohistochemical analysis of human prostate gland tissue (formalin/PFA-fixed paraffin-embedded section). Panel A : merged staining of anti-Collagen VI (ab271938, magenta; Opal™690), anti-Cytokeratin 5 (ab236216, green; Opal™520) and anti-Prostate Specific Antigen (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 ab271938 (1/500), ab236216 (1/400), and 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. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182744).

• mIHC

Lab

Multiplex immunohistochemistry - 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 ab93741 at 1/200 dilution (1.02 µg/mL) (B), Collagen VI with ab271938 at 1/500 dilution (2.084 μg/ml) (C) and Cytokeratin 14 with 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 ab236439 for 30 mins, ab93741 for 10 mins, and 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Collagen VI antibody [EPR17072] - BSA and Azide free (AB271938)

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) (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 ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and 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 ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 - ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182744).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17072] - BSA and Azide free (AB271938)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182744).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17072] - BSA and Azide free (AB271938)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182744).

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Collagen VI antibody [EPR17072] - BSA and Azide free (AB271938)

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

Panel A : Merged staining of Collagen VI (ab182744; green), anti-CD68 (ab213363; red) and anti-Lamin B1 (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), ab213363 (1/1000 dilution) and 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182744).

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Collagen VI antibody [EPR17072] - BSA and Azide free (AB271938)

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 (ab274418; gray; Opal™690) at 1 : 2000 (0.263 μg/ml) [Panel B], anti-CYP11A1 stained on adrenal cortex (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 ab274418, 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.  This data was developed using ab182744, the same antibody clone in a different buffer formulation.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Collagen VI antibody [EPR17072] - BSA and Azide free (AB271938)

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

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat ovary tissue staining AMH with ab272221 at a 1 : 2000 (0.26 μg/ml) dilution, ab182744 Collagen VI used at 1 : 2000 (0.76 ug/ml) dilution and ab270534 DDX4 / MVH used at 1 : 5000 (0.1 μg/ml) dilution.

Panel A : merged staining of anti-AMH (green; Opal™570), anti-Collagen VI (magenta; Opal™690) and anti-DDX4 / MVH (gray; Opal™570) on rat ovary.

Panel B : anti-AMH staining granulosa cells in rat ovary.

Panel C : ant-Collagen VI staining stroma in rat ovary.

Panel D : ant-DDX4 / MVH staining oocytes in rat ovary.

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

The section was incubated in three rounds of staining : in the order of ab272221, ab182744 and ab270534 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 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 VI antibody [EPR17072] - BSA and Azide free (AB271938)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182744).

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Collagen VI antibody [EPR17072] - BSA and Azide free (AB271938)

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

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse ovary tissue staining AMH with ab272221 at a 1 : 2000 (0.26 μg/ml) dilution, ab182744 Collagen VI used at 1 : 2000 (0.76 ug/ml) dilution and ab270534 DDX4 / MVH used at 1 : 5000 (0.1 μg/ml) dilution.

Panel A : merged staining of anti-AMH (green; Opal™570), anti-Collagen VI (magenta; Opal™690) and anti-DDX4 / MVH (gray; Opal™570) on mouse ovary.

Panel B : anti-AMH staining granulosa cells in mouse ovary.

Panel C : ant-Collagen VI staining stroma in mouse ovary.

Panel D : ant-DDX4 / MVH staining oocytes in mouse ovary.

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

The section was incubated in three rounds of staining : in the order of ab272221, ab182744 and ab270534 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Collagen VI antibody [EPR17072] - BSA and Azide free (AB271938)

Immunohistochemistry analysis of Formalin/PFA-fixed paraffin-embedded sections Mouse Prostate tissue labelling E Cadherin with ab324191 at 1 : 500 dilution (B), Cytokeratin 5 with ab236216 at 1 : 1500 dilution (C) and Collagen VI with ab271938 at 1 : 2000 dilution (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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-E Cadherin (green; Opal™690), anti-Cytokeratin 5 (magenta; Opal™520) and anti-Collagen VI (gray; Opal™570) on mouse prostate.

Panel B : anti-E Cadherin staining epithelium in mouse prostate.

Panel C : ant-Cytokeratin 5 staining basal cells in mouse prostate.

Panel D : ant-Collagen VI staining stoma in mouse prostate.

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

The section was incubated in three rounds of staining : in the order of ab324191, ab236216 and 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17072] - BSA and Azide free (AB271938)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182744).