Otx1 + Otx2 Recombinant Rabbit Monoclonal Antibody [PSH01-62]
cat.: HA721705
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Rat, Mouse |
| Applications: | WB, IHC-P, IF-Cell, FC |
| Clonality: | Monoclonal |
| Clone number: | PSH01-62 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| 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: 31.6 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human OTX2 aa 1-289. |
| Positive control: | 293T transfected with FLAG-tagged Otx2 cell lysate, rat eye tissue, 293T overexpress with Otx2. |
| Subcellular location: | Nucleus. |
| Recommended Dilutions:
WB IHC-P IF-Cell FC |
1:1,000 1:200 1:1,000 1:1,000 |
| Uniprot #: | SwissProt: P32242 Human | P32243 Human | P80205 Mouse | P80206 Mouse | Q63410 Rat | Q64201 Rat |
| Alternative names: | FLJ38361 Homeobox protein OTX 1 Homeobox protein OTX1 Homeobox protein OTX2 MCOPS 5 MCOPS5 MGC15736 MGC45000 Orthodenticle 1 Orthodenticle 2 Orthodenticle homeobox 1 Orthodenticle homeobox 2 Orthodenticle homolog 1 Orthodenticle homolog 2 (Drosophila) Orthodenticle homolog 2 Orthodenticle1 Orthodenticle2 Otx 1 Otx 2 otx1 OTX1_HUMAN otx2 OTX2_HUMAN |
Images
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Fig1:
Western blot analysis of Otx1 + Otx2 on different lysates with Rabbit anti-Otx1 + Otx2 antibody (HA721705) at 1/1,000 dilution. Lane 1: 293T transfected with FLAG-tagged empty control cell lysate Lane 2: 293T transfected with FLAG-tagged Otx2 cell lysate Lysates/proteins at 30 µg/Lane. Predicted band size: 32 kDa Observed band size: 32 kDa Exposure time: 24 seconds; 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 (HA721705) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Western blot analysis of Otx1 + Otx2 on different lysates with Rabbit anti-Otx1 + Otx2 antibody (HA721705) at 1/1,000 dilution. Lane 1: Human Otx2 recombinant protein, 50ng/Lane Lane 2: Human Otx1 recombinant protein, 50ng/Lane Exposure time: 5 seconds; 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 (HA721705) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature. |
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Fig3:
Immunohistochemical analysis of paraffin-embedded rat eye tissue with Rabbit anti-Otx1 + Otx2 antibody (HA721705) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA721705) 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. |
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Fig4:
Immunocytochemistry analysis of 293T overexpress with or without Otx2 cells labeling Otx1 + Otx2 with Rabbit anti-Otx1 + Otx2 antibody (HA721705) at 1/1,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Otx1 + Otx2 antibody (HA721705) 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 (M1305-2, 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. |
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Fig5:
Flow cytometric analysis of 293T overexpress with or without Otx2 cells labeling Otx1 + Otx2. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721705, 1μg/mL) (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). |
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