Osteopontin Recombinant Rabbit Monoclonal Antibody [PSH09-24]
cat.: HA723082
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse |
| Applications: | WB, IF-Cell, FC |
| Clonality: | Monoclonal |
| Clone number: | PSH09-24 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 35 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human Osteopontin aa 1-314. |
| Positive control: | RAW264.7 treated with 100ng/mL LPS for 4 hours add 1μg/mL BFA for last 3 hours cell lysate, A549 cell lysate, U-87 MG cell lysate, U-87 MG, A549. |
| Subcellular location: | Secreted. |
| Recommended Dilutions:
WB IF-Cell FC |
1:2,000 1:1,000-1:20,000 1:1,000 |
| Uniprot #: | SwissProt: P10451 Human | P10923 Mouse |
| Alternative names: | BNSP Bone sialoprotein 1 Bone sialoprotein I BSP I BSPI Early T lymphocyte activation 1 ETA 1 ETA1 MGC110940 Nephropontin OPN Osteopontin osteopontin/immunoglobulin alpha 1 heavy chain constant region fusion protein OSTP_HUMAN PSEC0156 secreted phosphoprotein 1 (osteopontin bone sialoprotein I early T lymphocyte activation 1) secreted phosphoprotein 1 (osteopontin, bone sialoprotein I, early T-lymphocyte activation 1) Secreted phosphoprotein 1 SPP 1 SPP-1 SPP1 SPP1/CALPHA1 fusion Urinary stone protein Uropontin |
Images
|
Fig1:
Western blot analysis of Osteopontin on different lysates with Rabbit anti-Osteopontin antibody (HA723082) at 1/2,000 dilution. Lane 1: RAW264.7 cell lysate Lane 2: RAW264.7 treated with 100ng/mL LPS for 4 hours, add 1μg/mL BFA for last 3 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 35 kDa Observed band size: 55 kDa Exposure time: 16 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 (HA723082) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
|
Fig2:
Western blot analysis of Osteopontin on different lysates with Rabbit anti-Osteopontin antibody (HA723082) at 1/2,000 dilution. Lane 1: A549 cell lysate Lane 2: U-87 MG cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 35 kDa Observed band size: 55 kDa Exposure time: 1 minute; ECL: K1802; 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 (HA723082) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
|
Fig3:
Immunocytochemistry analysis of U-87 MG cells labeling Osteopontin with Rabbit anti-Osteopontin antibody (HA723082) at 1/20,000 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-Osteopontin antibody (HA723082) at 1/20,000 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. |
|
Fig4:
Immunocytochemistry analysis of RAW264.7 cells treated with 100ng/mL LPS for 4 hours, add 1μg/mL BFA for last 3 hours labeling Osteopontin with Rabbit anti-Osteopontin antibody (HA723082) at 1/1,000 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-Osteopontin antibody (HA723082) at 1/1,000 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. |
|
Fig5:
Flow cytometric analysis of A549 cells labeling Osteopontin. Cells were fixed and permeabilized. Then stained with the primary antibody (HA723082, 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). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.