Anti-S100 alpha 2/S100A2 antibody [EPR5392]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-S100 alpha 2/S100A2 antibody [EPR5392] (AB109494)

Immunocytochemistry/ Immunofluorescence analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labeling S100 alpha 2 using ab109494. The cells were fixed with 100% Methanol then permeabilized with 0.1% Triton X-100. The cells were then incubated with ab109494 at 1 : 100 dilution followed by a further incubation with a Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI. Cells were counterstained using ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1 : 200 dilution (shown in red). Secondary antibody only control : PBS instead of the primary antibody.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 alpha 2/S100A2 antibody [EPR5392] (AB109494)

ab109494 at 1/250 dilution staining S100 alpha 2/S100A2 in paraffin-embedded Human squamous cell cervical carcinoma tissue

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-S100 alpha 2/S100A2 antibody [EPR5392] (AB109494)

Overlay histogram showing A431 cells stained with ab109494 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109494 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 alpha 2/S100A2 antibody [EPR5392] (AB109494)

Immunohistochemical analysis of Paraffin-embedded sections human skin tissue labelling S100 alpha 2 with ab109494 at 1/1000 dilution, followed by a ready to use secondary Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Staining on human skin tissue is observed. Counter stained with Haematoxylin. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 alpha 2/S100A2 antibody [EPR5392] (AB109494)

Immunohistochemical analysis of Paraffin-embedded sections human lung carcinoma tissue labelling S100 alpha 2 with ab109494 at 1/1000 dilution, followed by a ready to use secondary Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Staining on human lung carcinoma tissue is observed. Counter stained with Haematoxylin. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-S100 alpha 2/S100A2 antibody [EPR5392] (AB109494)

Intracellular Flow Cytometry analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labeling S100 alpha 2 with purified ab109494 at 1/20 dilution (10 ug/ml) (Red). Cells were fixed with 80% Methanol and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as a isotype control. Cell without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

• IP

Lab

Immunoprecipitation - Anti-S100 alpha 2/S100A2 antibody [EPR5392] (AB109494)

S100 alpha 2/S100A2 was immunoprecipitated from 0.35 mg HaCaT (Human skin keratinocyte) whole cell lysate 10 μg with ab109494 at 1/20 dilution (1μg). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HaCaT (Human skin keratinocyte) whole cell lysate 10 μg

Lane 2 : ab109494 IP in HaCaT whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab109494 in HaCaT whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-S100 alpha 2/S100A2 antibody [EPR5392] (ab109494)

Predicted band size: 11 kDa

Observed band size: 11 kDa

false

• WB

Lab

Western blot - Anti-S100 alpha 2/S100A2 antibody [EPR5392] (AB109494)

All lanes:

Western blot - Anti-S100 alpha 2/S100A2 antibody [EPR5392] (ab109494) at 1/1000 dilution

Lane 1:

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

Lane 2:

HaCaT (Human skin keratinocyte) whole cell lysate at 15 µg

Lane 3:

BxPC-3 (Human pancreas adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 4:

A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate at 15 µg

Secondary

All lanes:

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

Predicted band size: 11 kDa

Observed band size: 11 kDa

false