S100 alpha 6 Recombinant Rabbit Monoclonal Antibody [JF0976]
cat.: ET1702-28
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality: Monoclonal
Clone number: JF0976
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. 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: 10 kDa
Isotype: IgG
Immunogen: Recombinant full length protein of Human S100 alpha 6 aa 1-90 / 90.
Positive control: HeLa cell lysate, A549 cell lysate, Neuro-2a cell lysate, C6 cell lysate, mouse lung tissue lysate, C6, human kidney tissue, human stomach carcinoma tissue, human colon carcinoma tissue, mouse brain tissue, mouse kidney tissue, rat kidney tissue.
Subcellular location: Nucleus envelope, Cytoplasm, Cell membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
1:2,000-1:5,000
1:100-1:500
1:100-1:500
1:50-1:1,000
1:50-1:100
Use at an assay dependent concentration.
Uniprot #: SwissProt: P06703 Human | P14069 Mouse | P05964 Rat
Alternative names: 2A9 5B10 CABP CACY Calcyclin Growth factor inducible protein 2A9 Growth factor-inducible protein 2A9 MLN 4 MLN4 OTTHUMP00000015472 OTTHUMP00000015473 PRA PRAGrowth factor inducible protein 2A9 Prolactin receptor associated protein Prolactin receptor-associated protein Protein S100 A6 Protein S100-A6 S100 A6 S100 calcium binding protein A6 (calcyclin) S100 calcium binding protein A6 S100 calcium-binding protein A6 S100A6 S10A6_HUMAN
Images
ET1702-28_1.jpg Fig1: Western blot analysis of S100 alpha 6 on different lysates with Rabbit anti-S100 alpha 6 antibody (ET1702-28) at 1/2,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: A549 cell lysate
Lane 3: Neuro-2a cell lysate
Lane 4: C6 cell lysate
Lane 5: Mouse lung tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 10 kDa
Observed band size: 10 kDa

Exposure time: 10 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 (ET1702-28) 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.
ET1702-28_2.jpg Fig2: Immunocytochemistry analysis of C6 cells labeling S100 alpha 6 with Mouse anti-S100 alpha 6 antibody (ET1702-28) 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 Mouse anti-S100 alpha 6 antibody (ET1702-28) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution.
ET1702-28_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-S100 alpha 6 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1702-28, 1/50) 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.
ET1702-28_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue using anti-S100 alpha 6 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1702-28, 1/50) 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.
ET1702-28_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-S100 alpha 6 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1702-28, 1/50) 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.
ET1702-28_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-S100 alpha 6 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1702-28, 1/50) 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.
ET1702-28_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-S100 alpha 6 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1702-28, 1/50) 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.
ET1702-28_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-S100 alpha 6 antibody (ET1702-28) 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 (ET1702-28) 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.
ET1702-28_9.jpg Fig9: Flow cytometric analysis of S100 alpha 6 was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1702-28, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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