Fibroblast Marker (Vimentin, alpha smooth muscle Actin, Hsp47, S100A4) Antibody Panel - Human, Mouse

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Fibroblast Marker (Vimentin, alpha smooth muscle Actin, Hsp47, S100A4) Antibody Panel - Human, Mouse (AB254015)

ab92547 staining Vimentin in wild-type HAP1 cells (top panel) and VIM knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab92547 at 0.5μg/ml and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Fibroblast Marker (Vimentin, alpha smooth muscle Actin, Hsp47, S100A4) Antibody Panel - Human, Mouse (AB254015)

ab109117 staining Hsp47 in Human colon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/300). An undiluted HRP-conjugated mouse anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Fibroblast Marker (Vimentin, alpha smooth muscle Actin, Hsp47, S100A4) Antibody Panel - Human, Mouse (AB254015)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human smooth muscle tissue labeling alpha smooth muscle Actin with purified ab32575 at a dilution of 1/200. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500)

Negative control using PBS instead of primary antibody.

Counterstained with hematoxylin.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Fibroblast Marker (Vimentin, alpha smooth muscle Actin, Hsp47, S100A4) Antibody Panel - Human, Mouse (AB254015)

Immunocytochemistry/Immunofluorescence analysis of A431 (human epidermoid carcinoma) cells labeling alpha smooth muscle Actin (green) with purified ab32575 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counterstained with ab7291, anti-Tubulin (mouse mAb) at 1/1000 followed by ab150120 Alexa Fluor^®594 goat anti-mouse secondary (1/1000). Nuclei were counterstained with DAPI (blue).

For negative control 1, rabbit primary antibody and anti-mouse secondary antibody (ab150120) were used. For negative control 2, ab7291 (mouse primary antibody) was used followed by anti-rabbit secondary antibody (ab150077).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Fibroblast Marker (Vimentin, alpha smooth muscle Actin, Hsp47, S100A4) Antibody Panel - Human, Mouse (AB254015)

Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab92547 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Fibroblast Marker (Vimentin, alpha smooth muscle Actin, Hsp47, S100A4) Antibody Panel - Human, Mouse (AB254015)

Immunohistochemical analysis of paraffin-embedded Human gastric carcinoma tissue labeling S100A4 using ab197896 at 1/2000 dilution. Goat Anti-Rabbit IgG H&L (HRP) ab97051 was used as a secondary antibody at 1/500 dilution. Cells were counterstained with Hematoxylin.

Inset image : negative control obtained using PBS instead of ab197896 and secondary antibody only.
Note : Cytoplasm and nuclear staining on human gastric carcinoma tissue was observed.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Fibroblast Marker (Vimentin, alpha smooth muscle Actin, Hsp47, S100A4) Antibody Panel - Human, Mouse (AB254015)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse smooth muscle tissue labeling alpha smooth muscle Actin with purified ab32575 at a dilution of 1/200. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).

Negative control using PBS instead of primary antibody.

Counterstained with hematoxylin.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Fibroblast Marker (Vimentin, alpha smooth muscle Actin, Hsp47, S100A4) Antibody Panel - Human, Mouse (AB254015)

Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3(Mouse embryonic fibroblast) cells labeling alpha smooth muscle Actin with purified ab32575 at 1/500 dilution (5.2 μg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 μg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Fibroblast Marker (Vimentin, alpha smooth muscle Actin, Hsp47, S100A4) Antibody Panel - Human, Mouse (AB254015)

Immunohistochemical staining of paraffin embedded mouse kidney with purified ab92547 at a working dilution of 1/250. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• WB

Supplier Data

Western blot - Fibroblast Marker (Vimentin, alpha smooth muscle Actin, Hsp47, S100A4) Antibody Panel - Human, Mouse (AB254015)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : Hsp47 knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HeLa whole cell lysate (20 μg)
Lane 4 : HepG2 whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - Anti-Hsp47 antibody [EPR4217] (ab109117) observed at 46 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab109117 was shown to recognize Hsp47 in wild-type HAP1 cells as signal was lost at the expected MW in Hsp47 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Hsp47 knockout samples were subjected to SDS-PAGE. ab109117 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.

All lanes:

Western blot - Anti-Hsp47 antibody [EPR4217] (<a href='/en-us/products/primary-antibodies/hsp47-antibody-epr4217-ab109117'>ab109117</a>)

Predicted band size: 46 kDa

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