| Product Type: | Rabbit polyclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IHC-P, FC, IP |
| Clonality: | Polyclonal |
| 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: | 1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Immunogen affinity purified. |
| Molecular weight: | Predicted band size: 64 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human Metadherin aa 18-253. |
| Positive control: | 786-0 cell lysate, Raji cell lysate, Raji, N2A, human breast tissue, mouse brain tissue. |
| Subcellular location: | Endoplasmic reticulum membrane, Nucleus membrane, Cell junction, tight junction, Nucleus, nucleolus, Cytoplasm, perinuclear region. |
| Recommended Dilutions:
WB IF-Cell IHC-P FC IP |
1:2,000 1:50-1:200 1:100-1:5,000 1:50-1:200 1-2μg/sample |
| Uniprot #: | SwissProt: Q86UE4 Human | Q80WJ7 Mouse | Q9Z1W6 Rat |
| Alternative names: | 3D3 3D3/LYRIC AEG 1 AEG-1 AEG1 Astrocyte elevated gene 1 Astrocyte elevated gene-1 protein LYRIC LYRIC/3D3 LYRIC_HUMAN Lysine rich CEACAM1 associated protein Lysine rich CEACAM1 co isolated protein Lysine-rich CEACAM1 co-isolated protein Metadherin Metastasis adhesion protein MTDH Protein LYRIC |
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Fig1:
Western blot analysis of Metadherin / LYRIC on different lysates with Rabbit anti-Metadherin / LYRIC antibody (R1511-10) at 1/2,000 dilution. Lane 1: 786-0 cell lysate Lane 2: Raji cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 64 kDa Observed band size: 75 kDa Exposure time: 6 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 (R1511-10) 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. |
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Fig2:
Immunocytochemistry analysis of Raji cells labeling Metadherin / LYRIC with Rabbit anti-Metadherin / LYRIC antibody (R1511-10) 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-Metadherin / LYRIC antibody (R1511-10) 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. |
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Fig3: ICC staining Metadherin(LYRIC) in N2A cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue with Rabbit anti-Metadherin / LYRIC antibody (R1511-10) at 1/5,000 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 (R1511-10) at 1/5,000 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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Fig5:
Immunohistochemical analysis of paraffin-embedded rat small intestine tissue with Rabbit anti-Metadherin / LYRIC antibody (R1511-10) at 1/5,000 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 (R1511-10) at 1/5,000 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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Fig6: Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-Metadherin(LYRIC) antibody. Counter stained with hematoxylin. |
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Fig7:
Flow cytometric analysis of Raji cells labeling Metadherin / LYRIC. Cells were fixed and permeabilized. Then stained with the primary antibody (R1511-10, 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). |
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Fig8:
Metadherin / LYRIC was immunoprecipitated from 0.2 mg Raji cell lysate with R1511-10 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using R1511-10 at 1/2,000 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: Raji cell lysate (input) Lane 2: R1511-10 IP in Raji cell lysate Lane 3: Rabbit IgG instead of R1511-10 in Raji cell lysate Blocking/Dilution buffer: primary antibody dilution (K1803) Exposure time: 2 seconds; ECL: K1801 |