Anti-SS18 antibody [EPR23636-98]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SS18 antibody [EPR23636-98] (AB254405)

Immunohistochemical analysis of paraffin-embedded Human cervical carcinoma tissue labeling SS18 with ab254405 at 1/5000 (5.73 ug/ml) dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in human cervical carcinoma. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SS18 antibody [EPR23636-98] (AB254405)

Immunohistochemical analysis of paraffin-embedded Human synovial sarcoma tissue labeling SS18 with ab254405 at 1/5000 (5.73 ug/ml) dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in Human synovial sarcoma (PMID : 27120803). Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IP

Supplier Data

Immunoprecipitation - Anti-SS18 antibody [EPR23636-98] (AB254405)

SS18 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab254405 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254405 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug

Lane 2 : ab254405 IP in HeLa whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab254405 in HeLa whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 6 seconds.

Lysates were made freshly and used in WB test immediately to minimize protein degradation.

All lanes:

Immunoprecipitation - Anti-SS18 antibody [EPR23636-98] (ab254405)

Predicted band size: 46 kDa

Observed band size: 45 kDa,55 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SS18 antibody [EPR23636-98] (AB254405)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling SS18 with ab254405 at 1/5000 (5.73 ug/ml) dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in mouse colon. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SS18 antibody [EPR23636-98] (AB254405)

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling SS18 with ab254405 at 1/5000 (5.73 ug/ml) dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in rat cardiac muscle. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• WB

Lab

Western blot - Anti-SS18 antibody [EPR23636-98] (AB254405)

False colour image of Western blot : Anti-SS18 antibody [EPR23636-98] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab254405 was shown to bind specifically to SS18. A band was observed at 51/56 kDa in wild-type HeLa cell lysates with no signal observed at this size in SS18 knockout cell line ab265493 (knockout cell lysate ab259156). To generate this image, wild-type and SS18 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-SS18 antibody [EPR23636-98] (ab254405) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SS18 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human SS18 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ss18-knockout-hela-cell-line-ab265493'>ab265493</a>)

Lane 3:

HEK-293 cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Predicted band size: 46 kDa

Observed band size: 51 kDa,56 kDa

false

• WB

Lab

Western blot - Anti-SS18 antibody [EPR23636-98] (AB254405)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID : 23540691;29374058)

Lysates were made freshly and used in WB test immediately to minimize protein degradation.

Exposure times : Lanes 1-3 : 59 seconds; Lane 4 : 3 minutes; Lane 5 : 59 seconds.

All lanes:

Western blot - Anti-SS18 antibody [EPR23636-98] (ab254405) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

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

Lane 4:

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

Lane 5:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 46 kDa

Observed band size: 45 kDa,55 kDa

false