Anti-S100 Calcium Binding Protein A13/S100A13 antibody [EPR4510]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 Calcium Binding Protein A13/S100A13 antibody [EPR4510] (AB109252)

ab109252, at a 1/250 dilution, staining S100 Calcium Binding Protein A13/S100A13 in Human muscle, by Immunohistochemistry, Paraffin-embedded tissue.

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

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-S100 Calcium Binding Protein A13/S100A13 antibody [EPR4510] (AB109252)

Immunocytochemistry/ Immunofluorescence analysis of HUVEC (human umbilical vein endothelial cell) cells labeling S100 Calcium Binding Protein A13/S100A13 with purified ab109252 at 1/100 dilution (10 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-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 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/mL) was used as the secondary antibody only control.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-S100 Calcium Binding Protein A13/S100A13 antibody [EPR4510] (AB109252)

Intracellular Flow Cytometry analysis of HUVEC (Human umbilical vein endothelial cell) cells labeling S100 Calcium Binding Protein A13/S100A13 with purified ab109252 at 1/100 dilution (10 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cells without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 Calcium Binding Protein A13/S100A13 antibody [EPR4510] (AB109252)

ab109252, at a 1/250 dilution, staining S100 Calcium Binding Protein A13/S100A13 in Human brain, by Immunohistochemistry, Paraffin-embedded tissue.

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

• IP

Lab

Immunoprecipitation - Anti-S100 Calcium Binding Protein A13/S100A13 antibody [EPR4510] (AB109252)

Purified ab109252 at 1/50 dilution (2μg) immunoprecipitating S100 Calcium Binding Protein A13/S100A13 in Human heart lysate.
Lane 1 (input) : Human heart lysate 10μg
Lane 2 (+) : ab109252 + Human heart lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab109252 in Human heart lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 11 kDa

All lanes:

Immunoprecipitation - Anti-S100 Calcium Binding Protein A13/S100A13 antibody [EPR4510] (ab109252)

Predicted band size: 11 kDa

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• WB

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Western blot - Anti-S100 Calcium Binding Protein A13/S100A13 antibody [EPR4510] (AB109252)

All lanes:

Western blot - Anti-S100 Calcium Binding Protein A13/S100A13 antibody [EPR4510] (ab109252) at 1/1000 dilution

Lane 1:

Human thyroid tissue lysate at 10 µg

Lane 2:

Human testis tissue lysate at 10 µg

Lane 3:

HUVEC lysate at 10 µg

Lane 4:

ECV304 lysate at 10 µg

Predicted band size: 11 kDa

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• WB

Lab

Western blot - Anti-S100 Calcium Binding Protein A13/S100A13 antibody [EPR4510] (AB109252)

Lanes 1 - 4 : Merged signal (red and green). Green - ab109252 observed at 13 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab109252 was shown to react with S100 Calcium Binding Protein A13/S100A13 in wild-type HEK-293T cells in Western blot with loss of signal observed in S100A13 knockout cell line ab266065 (S100A13 knockout cell lysate ab258183). Wild-type HEK-293T and S100A13 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab109252 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-S100 Calcium Binding Protein A13/S100A13 antibody [EPR4510] (ab109252) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

S100A13 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human S100A13 (S100 Calcium Binding Protein A13) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-s100a13-s100-calcium-binding-protein-a13-knockout-hek-293t-cell-line-ab266065'>ab266065</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Predicted band size: 11 kDa

Observed band size: 13 kDa

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