All lanes : Anti-IGFBP2 antibody [EPR18012-257] (ab188200) at 1/1000 dilution

Lane 1 : Human serum
Lane 2 : Human liver lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

Developed using the ECL technique.

Predicted band size: 33 kDa
Observed band size: 33 kDa


Exposure time: 3 minutes

Blocking: 5% NFDM/TBST.

The band in lane 1 is human IgG heavy chain which is often observed in serum and plasma samples.

Immunohistochemical analysis of paraffin-embedded mouse choroid plexus tissue labeling IGFBP2 with ab188200 at 1/1000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining on mouse choroid plexus (PMID: 7525264; PMID: 7678219) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a Goat Anti-Rabbit IgG H&L (HRP) ready to use.

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

Immunohistochemical analysis of 4% PFA fixed, 0.2% TritonX-100 permeabilized mouse brain (choroid plexus) tissue labeling IGFBP2 with ab188200 at 1/500 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution. Cytoplasmic staining in the epithelial cells of choroid plexus on mouse tissue section is observed. Counter stained with DAPI.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.
Perform heat mediated antigen retrieval using Tris-EDTA (pH 9.0) (ab94681).

Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized T-47D (human ductal breast epithelial tumor epithelial cell) cell line labeling IGFBP2 with ab188200 at 1/60 (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

IGFBP2 was immunoprecipitated from 0.35 mg of human serum with ab188200 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab188200 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: Human serum  10 μg (Input).
Lane 2: ab188200 IP in Human serum (+).
Lane 3: Rabbit monoclonal IgG (ab172730) instead of  ab188200 in human serum (-).

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

Exposure time: 1 second.

The band in lane 1 is human IgG heavy chain which is often observed in serum and plasma samples.

All lanes : Anti-IGFBP2 antibody [EPR18012-257] (ab188200) at 1/1000 dilution

Lane 1 : Mouse serum
Lane 2 : T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate
Lane 3 : Rat serum
Lane 4 : RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 33 kDa
Observed band size: 33 kDa


Exposure time: 3 minutes

Blocking: 5% NFDM/TBST.

The expression profile observed on T-47D is consistent with what has been described in the literature (PMID: 23515291).

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling IGFBP2 with ab188200 at 1/1000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining on mouse liver (PMID: 7678219) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a Goat Anti-Rabbit IgG H&L (HRP) ready to use.

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

Immunohistochemical analysis of 4% PFA fixed, 0.2% TritonX-100 permeabilized rat brain (choroid plexus) tissue labeling IGFBP2 with ab188200 at 1/500 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution. Cytoplasmic staining in the epithelial cells of choroid plexus on rat tissue section is observed. Counter stained with DAPI.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.
Perform heat mediated antigen retrieval using Tris-EDTA (pH 9.0) (ab94681).

Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cell line labeling IGFBP2 with ab188200 at 1/60 (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.