WB
Western blot analysis of Metadherin / LYRIC on different lysates with Rabbit anti-Metadherin / LYRIC antibody at 1/2,000 dilution. Lane 1: 786-0 cell lysate, Lane 2: Raji cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 6 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue with Rabbit anti-Metadherin / LYRIC antibody 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 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.IHC
Immunohistochemical analysis of paraffin-embedded rat small intestine tissue with Rabbit anti-Metadherin / LYRIC antibody 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 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.ICC/IF
Immunocytochemistry analysis of Raji cells labeling Metadherin / LYRIC with Rabbit anti-Metadherin / LYRIC antibody 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of Raji cells labeling Metadherin / LYRIC. Cells were fixed and permeabilized. Then stained with the primary antibody (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
Metadherin / LYRIC was immunoprecipitated from 0.2 mg Raji cell lysate with Rabbit anti-Metadherin / LYRIC antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-Metadherin / LYRIC antibody 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: Rabbit anti-Metadherin / LYRIC antibody IP in Raji cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Metadherin / LYRIC antibody in Raji cell lysate, Blocking/Dilution buffer: primary antibody dilution, Exposure time: 2 seconds.| Product Name | Metadherin / LYRIC Rabbit Polyclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | MTDH (metastasis adhesion protein), also known as astrocyte elevated gene-1 protein (AEG1) or lysine-rich CEACAM1 co-isolated protein (LYRIC), is a 582 amino acid single-pass membrane protein. Overexpressed in various cancers, MTDH activates the transcription factor NFκB and promotes anchorage-independent growth of immortalized astrocytes and melanocytes, which are key components of tumor growth. MTDH is also thought to enhance the seeding of tumor cells to the target organ endothelium. Localized mainly to the endoplasmic reticulum membrane, MTDH also translocates from the cytoplasm to the nucleus when induced by TNFα. MTDH also localizes to tight junctions (TJ) during the maturation of TJ complexes in epithelial cells. MTDH is found at highest levels in heart, skeletal muscle, tongue and small intestine. |
| Immunogen | Recombinant protein within human Metadherin aa 18-253. |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:2000 IHC:1:100-1:5000 ICC/IF:1:50-1:200 FC:1:50-1:200 IP:1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | 3D3 antibody 3D3/LYRIC antibody AEG 1 antibody AEG-1 antibody AEG1 antibody Astrocyte elevated gene 1 antibody Astrocyte elevated gene-1 protein antibody LYRIC antibody LYRIC/3D3 antibody LYRIC_HUMAN antibody Lysine rich CEACAM1 associated protein antibody Lysine rich CEACAM1 co isolated protein antibody Lysine-rich CEACAM1 co-isolated protein antibody Metadherin antibody Metastasis adhesion protein antibody MTDH antibody Protein LYRIC antibody |
|---|---|
| Molecular Weight(MW) | 64kDa(Observed band size: 75kDa) |
| Cellular Localization | Endoplasmic reticulum membrane, Nucleus membrane, Cell junction, tight junction, Nucleus, nucleolus, Cytoplasm, perinuclear region. |
| SwissProt ID | Q86UE4 |
|---|
WB
Western blot analysis of Metadherin / LYRIC on different lysates with Rabbit anti-Metadherin / LYRIC antibody at 1/2,000 dilution. Lane 1: 786-0 cell lysate, Lane 2: Raji cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 6 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue with Rabbit anti-Metadherin / LYRIC antibody 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 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat small intestine tissue with Rabbit anti-Metadherin / LYRIC antibody 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 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.
ICC/IF
Immunocytochemistry analysis of Raji cells labeling Metadherin / LYRIC with Rabbit anti-Metadherin / LYRIC antibody 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of Raji cells labeling Metadherin / LYRIC. Cells were fixed and permeabilized. Then stained with the primary antibody (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
Metadherin / LYRIC was immunoprecipitated from 0.2 mg Raji cell lysate with Rabbit anti-Metadherin / LYRIC antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-Metadherin / LYRIC antibody 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: Rabbit anti-Metadherin / LYRIC antibody IP in Raji cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Metadherin / LYRIC antibody in Raji cell lysate, Blocking/Dilution buffer: primary antibody dilution, Exposure time: 2 seconds.| Application Notes | WB:1:2000 IHC:1:100-1:5000 ICC/IF:1:50-1:200 FC:1:50-1:200 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
