重组Anti-S100 Calcium Binding蛋白A13/S100A13抗体[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 brain, 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

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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.

• 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

Lab

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

This blot was produced using 14% SDS-PAGE containing 15 μg of HUVEC whole cell lysate per lane at 150V for 50 minutes before being transferred onto a 0.45 μm PVDF membrane at 72V for 50 minutes. The membrane was then blocked for 1hr using 5% NFDM/TBST, then incubated with ab109252 (1/1000) at room temperature for 1hr. After being washed three times in TBST, the membrane was incubated with Peroxidase conjugated goat anti-rabbit IgG at 1/2000 dilution for 1hr at room temperature. The membrane was washed three times again. Then the signal was developed using the ECL technique.

ab109252 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in WB. The image shows the band intensity remains relatively constant across all storage temperatures, demonstrating that antibody activity is not affected under these conditions. This data was generated as part of a structured antibody stability study supporting Abcam’s transition from cold‑chain to ambient shipping.

All lanes:

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

All lanes:

HUVEC whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/2000 dilution

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Exposure time: 7s

• 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='/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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• 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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