Anti-PIP2 antibody [2C11]

• ICC/IF

PubMed

Immunocytochemistry/ Immunofluorescence - Anti-PIP2 antibody [2C11] (AB11039)

Immunofluorescence analysis of Human SaOS-2 (Human osteosarcoma) cells, staining PIP2 with ab11039. Cells were either unstimulated (upper panel) or stimulated with direct current (lower panel).

Cells were fixed in formaldehyde, permeabilized and then blocked with 1% BSA for 20 min. Cells were then incubated with a primary antibody (1/200) overnight at 4°C. A FITC-conjugated anti-mouse IgG was used as the secondary antibody.

Image from Ozkucur N et al., BMC Cell Biol. 2011 Jan 22;12:4. Fig S4.; doi:10.1186/1471-2121-12-4; 22 January, 2011, BMC Cell Biology 2011, 12:4

• ICC

AbReview79509****

Immunocytochemistry - Anti-PIP2 antibody [2C11] (AB11039)

Immunocytochemistry analysis of methanol-fixed 0.3% tritonX-100 permeabilized Bone Osteosarcoma Epithelial Cells staining with ab11039 at 1/200. Secondary antibody was Cy3® anti-mouse at 1/300 dilution. Samples were incubated with the primary antibody for 16 hours at 4°C. Blocking was done using 5% serum for 1 hour at 25°C.

This image is courtesy of an anonymous Abreview

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PIP2 antibody [2C11] (AB11039)

IHC image of PIP2 staining in Human Normal Kidney formalin fixed paraffin embedded tissue section*.

Performed on a Leica BOND^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 20mins. The section was then incubated with ab11039 at 5μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

Although PIP2 is typically known as a signaling lipid located at the plasma membrane, recent researches has shown that it also plays important roles within the nucleus (PMID : 39621780, PMID : 31261688).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PIP2 antibody [2C11] (AB11039)

IHC image of PIP2 staining in Mouse Normal brain formalin fixed paraffin embedded tissue section.

Performed on a Leica BOND^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 20mins. The section was then incubated with ab11039 at 5μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Although PIP2 is typically known as a signaling lipid located at the plasma membrane, recent researches has shown that it also plays important roles within the nucleus (PMID : 39621780, PMID : 31261688).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-PIP2 antibody [2C11] (AB11039)

ICC/IF image of ab11039 stained human HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab11039, 5 μg/ml) overnight at +4°C.

The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgM (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa, Hek293 and MCF7 cells.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PIP2 antibody [2C11] (AB11039)

IHC image of PIP2 staining in mouse normal brain formalin fixed paraffin embedded tissue section. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab11039, 1μg/ml overnight at +4°C. An HRP-conjugated secondary (ab98679, 1/1000 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PIP2 antibody [2C11] (AB11039)

IHC image of PIP2 staining in a formalin fixed, paraffin embedded human normal kidney tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab11039, 1μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre