IGFBP2 Recombinant Rabbit Monoclonal Antibody [PSH21-02]

☑ Relative expression (RE)

Western blot analysis of IGFBP2 on different lysates with Rabbit anti-IGFBP2 antibody (HA724197) at 1/5,000 dilution.

Lane 1: SH-SY5Y cell lysate
Lane 2: T-47D cell lysate
Lane 3: MCF7 cell lysate
Lane 4: BxPC-3 cell lysate
Lane 5: JEG-3 cell lysate (low expression)
Lane 6: MEF cell lysate
Lane 7: COS-1 cell lysate
Lane 8: C2C12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 35 kDa
Observed band size: 35 kDa

Exposure time: Lane 1-6: 3 minutes; ECL: K1802;
Exposure time: Lane 7-8: 25 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA724197) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry analysis of SH-SY5Y cells labeling IGFBP2 with Rabbit anti-IGFBP2 antibody (HA724197) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-IGFBP2 antibody (HA724197) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Immunocytochemistry analysis of C2C12 cells labeling IGFBP2 with Rabbit anti-IGFBP2 antibody (HA724197) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-IGFBP2 antibody (HA724197) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Application: IHC-Fr

Species: Mouse

Site: brain (choroid plexus)

Sample: Frozen section

Antibody concentration: 1/500

Antigen retrieval: Not required

Application: IHC-Fr

Species: Rat

Site: brain (choroid plexus)

Sample: Frozen section

Antibody concentration: 1/500

Antigen retrieval: Not required

Immunohistochemical analysis of paraffin-embedded mouse choroid plexus tissue with Rabbit anti-IGFBP2 antibody (HA724197) at 1/50 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA724197) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-IGFBP2 antibody (HA724197) at 1/50 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA724197) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-IGFBP2 antibody (HA724197) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA724197) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Flow cytometric analysis of SH-SY5Y cells labeling IGFBP2.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA724197, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Flow cytometric analysis of C2C12 cells labeling IGFBP2.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA724197, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

IGFBP2 was immunoprecipitated from 0.2 mg C2C12 cell lysate with HA724197 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724197 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: C2C12 cell lysate (input)
Lane 2: HA724197 IP in C2C12 cell lysate
Lane 3: Rabbit IgG instead of HA724197 in C2C12 cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 19 seconds; ECL: K1801