SAM68 Recombinant Rabbit Monoclonal Antibody [JE47-15]
cat.: ET7109-52
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P
Clonality: Monoclonal
Clone number: JE47-15
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 48 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human SAM68 aa 1-108 / 443.
Positive control: HeLa cell lysate, A431 cell lysate, HepG2 cell lysate, HT-29 cell lysate, NIH/3T3 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, HeLa, NIH/3T3, human liver cancer tissue, human colon cancer tissue, human stomach cancer tissue, human brain tissue, human kidney tissue, mouse brain tissue, mouse kidney tissue, rat brain tissue, rat kidney tissue.
Subcellular location: Cytoplasm. Membrane. Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
1:5,000
1:100
1:1,000
Uniprot #: SwissProt: Q07666 Human | Q3U8T3 Mouse
Alternative names: FLJ34027 GAP associated tyrosine phosphoprotein p62 GAP-associated tyrosine phosphoprotein p62 KH domain containing RNA binding signal transduction associated 1 KH domain-containing KHDR1_HUMAN Khdrbs1 p21 Ras GTPase activating protein associated p62 p21 Ras GTPase-activating protein-associated p62 p62 p68 RNA-binding Sam68 signal transduction-associated protein 1 Src associated in mitosis 68 kDa protein Src-associated in mitosis 68 kDa protein
Images
ET7109-52_1.jpg Fig1: Western blot analysis of SAM68 on different lysates with Rabbit anti-SAM68 antibody (ET7109-52) at 1/5,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: A431 cell lysate (20 µg/Lane)
Lane 3: HepG2 cell lysate (20 µg/Lane)
Lane 4: HT-29 cell lysate (20 µg/Lane)
Lane 5: NIH/3T3 cell lysate (20 µg/Lane)
Lane 6: Mouse brain tissue lysate (40 µg/Lane)
Lane 7: Rat brain tissue lysate (40 µg/Lane)

Predicted band size: 48 kDa
Observed band size: 68 kDa

Exposure time: 3 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 (ET7109-52) 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.
ET7109-52_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling SAM68 with Rabbit anti-SAM68 antibody (ET7109-52) 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-SAM68 antibody (ET7109-52) 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.
ET7109-52_3.jpg Fig3: Immunocytochemistry analysis of NIH/3T3 cells labeling SAM68 with Rabbit anti-SAM68 antibody (ET7109-52) 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-SAM68 antibody (ET7109-52) 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.
ET7109-52_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human liver cancer tissue using anti-SAM68 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-52, 1/1,000) for 30 minutes 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.
ET7109-52_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-SAM68 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-52, 1/1,000) for 30 minutes 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.
ET7109-52_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue using anti-SAM68 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-52, 1/1,000) for 30 minutes 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.
ET7109-52_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-SAM68 antibody (ET7109-52) at 1/1,000 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 (ET7109-52) at 1/1,000 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.
ET7109-52_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-SAM68 antibody (ET7109-52) at 1/1,000 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 (ET7109-52) at 1/1,000 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.
ET7109-52_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-SAM68 antibody (ET7109-52) at 1/1,000 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 (ET7109-52) at 1/1,000 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.
ET7109-52_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-SAM68 antibody (ET7109-52) at 1/1,000 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 (ET7109-52) at 1/1,000 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.
ET7109-52_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-SAM68 antibody (ET7109-52) at 1/1,000 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 (ET7109-52) at 1/1,000 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.
ET7109-52_12.jpg Fig12: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-SAM68 antibody (ET7109-52) at 1/1,000 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 (ET7109-52) at 1/1,000 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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