Anti-S100A4 antibody [EPR2761(2)]

• IHC-P

AbReview33988****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR2761(2)] (AB124805)

Immunohistochemical analysis of Human lung tissue, staining S100A4 with unpurified ab124805.

Tissue was fixed with HOPE and blocked with blocking solution for 5 minutes at 25°C. Samples were incubated with primary antibody (1/1000 in diluent) for 1 hour at 25°C. An undiluted HRP-conjugated goat anti-rabbit polyclonal IgG was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR2761(2)] (AB124805)

Unpurified ab124805, at 1/250 dilution, staining S100A4 in paraffin-embedded Human tonsil tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-S100A4 antibody [EPR2761(2)] (AB124805)

ab124805 staining S100A4 in Jurkat (human acute T cell leukemia) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at 1/500. ab7291 and ab150120 were used as counterstains for primary antibody ab124805 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

Negative control 1 : Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
Negative control 2 : Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-S100A4 antibody [EPR2761(2)] (AB124805)

ab124805 staining S100A4 in HeLa (human cervix adenocarcinoma) by intracellular flow cytometry. Cells were fixed with 80% methanoland permeabilised with 0.1% Triton X-100 (in PBS). The sample was incubated with the primary antibody at a dilution of 1/80. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control : ab172730 rabbit monoclonal IgG (Black)

Unlabelled control : Cell without incubation with primary antibody and secondary antibody (Blue)

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR2761(2)] (AB124805)

ab124805 staining S100A4 in human tonsil tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1 : PBS in place of primary antibody.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-S100A4 antibody [EPR2761(2)] (AB124805)

ab124805 staining S100A4 in HeLa (human cervix adenocarcinoma) by intracellular flow cytometry. Cells were fixed with 80% methanoland permeabilised with 0.1% Triton X-100 (in PBS). The sample was incubated with the primary antibody at a dilution of 1/800. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control : ab172730 rabbit monoclonal IgG (Black)

Unlabelled control : Cell without incubation with primary antibody and secondary antibody (Blue)

• ICC/IF

AbReview35444****

Immunocytochemistry/ Immunofluorescence - Anti-S100A4 antibody [EPR2761(2)] (AB124805)

Unpurified ab124805 staining S100A4 in the A549 cell line from Human lungs by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with Triton X-100 0.25% in PBS. Samples were incubated with primary antibody (1/100) for 45 minutes at 25°C. A TRITC-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

• IP

Lab

Immunoprecipitation - Anti-S100A4 antibody [EPR2761(2)] (AB124805)

ab124805 immunoprecipitating S100A4. 10μg of cell lysate was incubated with primary antibody at a dilution of 1/30 and Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at a dilution of 1/1500.

Lane 1 : Human tonsil whole cell lysate (10ug)
Lane 2 : Human tonsil whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab124805 in human tonsil whole cell lysate

All lanes:

Immunoprecipitation - Anti-S100A4 antibody [EPR2761(2)] (ab124805)

Predicted band size: 12 kDa

Observed band size: 12 kDa

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• WB

Lab

Western blot - Anti-S100A4 antibody [EPR2761(2)] (AB124805)

Lanes 1 - 4 : Merged signal (red and green). Green - ab124805 observed at 12 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab124805 was shown to react with S100A4 in wild-type A549 cells in Western blot with loss of signal observed in S100A4 knockout cell line ab261865 (knockout cell lysate ab261674). Wild-type A549 and S100A4 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab124805 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-S100A4 antibody [EPR2761(2)] (ab124805) at 1/1000 dilution

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

S100A4 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human S100A4 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-s100a4-knockout-a549-cell-line-ab261865'>ab261865</a>)

Lane 3:

A-375 (Human malignant melanoma cell line) whole cell lysate at 20 µg

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Predicted band size: 12 kDa

Observed band size: 12 kDa

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• WB

Lab

Western blot - Anti-S100A4 antibody [EPR2761(2)] (AB124805)

Lanes 1 - 2 : Merged signal (red and green). Green - ab124805 observed at 11 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab124805 was shown to react with S100A4 in wild-type HeLa cells in Western blot with loss of signal observed in S100A4 knockout cell line ab265709 (S100A4 knockout cell lysate ab257046). Wild-type HeLa and S100A4 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab124805 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-S100A4 antibody [EPR2761(2)] (ab124805) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

S100A4 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human S100A4 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-s100a4-knockout-hela-cell-line-ab265709'>ab265709</a>)

Predicted band size: 12 kDa

Observed band size: 11 kDa

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• WB

Lab

Western blot - Anti-S100A4 antibody [EPR2761(2)] (AB124805)

Blocking Buffer : 5% NFDM /TBST

Diluting Buffer : 5% NFDM /TBST

All lanes:

Western blot - Anti-S100A4 antibody [EPR2761(2)] (ab124805) at 1/2500 dilution

Lane 1:

Human tonsil tissue lysate at 10 µg

Lane 2:

A375 (human malignant melanoma) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 12 kDa

Observed band size: 12 kDa

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• WB

Lab

Western blot - Anti-S100A4 antibody [EPR2761(2)] (AB124805)

Blocking buffer : 5% NFDM /TBST

Diluting Buffer : 5% NFDM /TBST

All lanes:

Western blot - Anti-S100A4 antibody [EPR2761(2)] (ab124805)

All lanes:

A549 (human lung carcinoma) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 12 kDa

Observed band size: 12 kDa

false

• WB

Unknown

Western blot - Anti-S100A4 antibody [EPR2761(2)] (AB124805)

All lanes:

Western blot - Anti-S100A4 antibody [EPR2761(2)] (ab124805) at 1/1000 dilution

Lane 1:

Human tonsil lysate at 10 µg

Lane 2:

A549 cell lysate at 10 µg

Lane 3:

A375 cell lysate at 10 µg

Lane 4:

Human small intestine lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 12 kDa

Observed band size: 12 kDa

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