Anti-Vimentin antibody [EPR3776] - Chicken IgY (Chimeric) - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [EPR3776] - Chicken IgY (Chimeric) - BSA and Azide free (AB316215)

This data was developed using ab316214, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized VIM knock out HeLa (VIM knockout human cervical adenocarcinoma epithelial cell line) cells labelling Vimentin with ab316214 at 1/5000 (0.16 ug/ml) dilution, followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in wild-type HeLa cell line (shown in green), and negative staining in VIM knock out HeLa cell line. The counterstain was observed in red. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [EPR3776] - Chicken IgY (Chimeric) - BSA and Azide free (AB316215)

This data was developed using ab316214, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling Vimentin with ab316214 at 1/5000 (0.161 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human lung.
The section was incubated with ab316214 for 30 mins at room temperature, followed by anti-chicken IgY antibody (ab97136) for 8 mins during the LeicaDS9800 kit staining procedure.
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-Vimentin antibody [EPR3776] - Chicken IgY (Chimeric) - BSA and Azide free (AB316215)

This data was developed using ab316214, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A) Wild-type HeLa (human cervix adenocarcinoma epithelial cell) cell pellet (B) VIM knockout HeLa (ab255446) cell pellet tissue labeling Vimentin with ab316214 at 1/20000 (0.04 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) Wild-type HeLa cell pellet, no staining on (B) VIM knockout HeLa (ab255446) cell pellet.
The section was incubated with ab316214 for 30 mins at room temperature, followed by anti-chicken IgY antibody (ab97136) for 8 mins during the LeicaDS9800 kit staining procedure.
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-Vimentin antibody [EPR3776] - Chicken IgY (Chimeric) - BSA and Azide free (AB316215)

This data was developed using ab316214, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling Vimentin with ab316214 at 1/5000 (0.161 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on interstitial cells of human breast cancer.
The section was incubated with ab316214 for 30 mins at room temperature, followed by anti-chicken IgY antibody (ab97136) for 8 mins during the LeicaDS9800 kit staining procedure.
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-Vimentin antibody [EPR3776] - Chicken IgY (Chimeric) - BSA and Azide free (AB316215)

This data was developed using ab316214, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Vimentin with ab316214 at 1/5000 (0.161 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on interstitial cells of human colon.
The section was incubated with ab316214 for 30 mins at room temperature, followed by anti-chicken IgY antibody (ab97136) for 8 mins during the LeicaDS9800 kit staining procedure.
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-Vimentin antibody [EPR3776] - Chicken IgY (Chimeric) - BSA and Azide free (AB316215)

This data was developed using ab316214, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Vimentin with ab316214 at 1/5000 (0.161 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on interstitial cells of rat colon.
The section was incubated with ab316214 for 30 mins at room temperature, followed by anti-chicken IgY antibody (ab97136) for 8 mins during the LeicaDS9800 kit staining procedure.
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-Vimentin antibody [EPR3776] - Chicken IgY (Chimeric) - BSA and Azide free (AB316215)

This data was developed using ab316214, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Vimentin with ab316214 at 1/5000 (0.161 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on interstitial cells of mouse colon.
The section was incubated with ab316214 for 30 mins at room temperature, followed by anti-chicken IgY antibody (ab97136) for 8 mins during the LeicaDS9800 kit staining procedure.
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.

• WB

Supplier Data

Western blot - Anti-Vimentin antibody [EPR3776] - Chicken IgY (Chimeric) - BSA and Azide free (AB316215)

This data was developed using ab316214, the same antibody clone in a different buffer formulation.

Negative control : Daudi (PMID : 17606622).

In Western blot, ab316214 was shown to bind specifically to VIM (Vimentin). Target of interest was observed at 55 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in VIM (Vimentin) knockout cell line (lane 2, knockout cell line ab255446/ knockout cell lysate ab263775).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-4 : 8 seconds, Lanes 5 : 3 seconds, Lanes 6 : 10 seconds

All lanes:

Western blot - Anti-Vimentin antibody [EPR3776] - Chicken IgY (Chimeric) (<a href='/en-us/products/primary-antibodies/vimentin-antibody-epr3776-chicken-igy-chimeric-ab316214'>ab316214</a>) at 1/1000 dilution

Lane 1:

parental HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

VIM (Vimentin) knockout HeLa whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

Daudi (human Burkitts lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 5:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 6:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Chicken IgY H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-chicken-igy-h-l-hrp-ab6877'>ab6877</a>) at 1/100000 dilution

Observed band size: 55 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-Vimentin antibody [EPR3776] - Chicken IgY (Chimeric) - BSA and Azide free (AB316215)

This data was developed using ab316214, the same antibody clone in a different buffer formulation.

The identity of higher MW band at approximately 100 kDa (in lane 1) is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Vimentin antibody [EPR3776] - Chicken IgY (Chimeric) (<a href='/en-us/products/primary-antibodies/vimentin-antibody-epr3776-chicken-igy-chimeric-ab316214'>ab316214</a>) at 1/1000 dilution

Lane 1:

Human lung tissue lysate at 20 µg

Lane 2:

Mouse lung tissue lysate at 20 µg

Lane 3:

Rat lung tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Chicken IgY H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-chicken-igy-h-l-hrp-ab6877'>ab6877</a>) at 1/100000 dilution

Observed band size: 55 kDa,36 kDa

false

Exposure time: 26s