VPS28 Recombinant Rabbit Monoclonal Antibody [JE55-08]
cat.: ET7110-94
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: JE55-08
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: 25 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human VPS28 aa 100-221 / 221.
Positive control: Mouse testis tissue lysate, human small intestine tissue lysate, rat bone marrow tissue lysate, N2A, human colon tissue, human prostate carcinoma tissue, human uterus tissue, mouse testis tissue, SH-SY5Y.
Subcellular location: Late endosome membrane, Cell membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:500-1:1,000
1:50-1:200
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: Q9UK41 Human | Q9D1C8 Mouse | B5DEN9 Rat
Alternative names: ESCRT-I complex subunit VPS28 H-Vps28 Vacuolar protein sorting 28 Vacuolar protein sorting 28 (yeast) Vacuolar protein sorting 28 homolog (S. cerevisiae) Vacuolar protein sorting 28 homolog Vacuolar protein sorting-associated protein 28 homolog VPS 28 Vps28 VPS28_HUMAN Yeast class E protein Vps28p homolog
Images
ET7110-94_1.jpg Fig1: Western blot analysis of VPS28 on different lysates with Rabbit anti-VPS28 antibody (ET7110-94) at 1/500 dilution.

Lane 1: Mouse testis tissue lysate
Lane 2: Human small intestine tissue lysate
Lane 2: Rat bone marrow tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 25 kDa
Observed band size: 25 kDa

Exposure time: 2 minutes;

12% 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 (ET7110-94) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
ET7110-94_2.jpg Fig2: Immunocytochemistry analysis of N2A cells labeling VPS28 with Rabbit anti-VPS28 antibody (ET7110-94) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-VPS28 antibody (ET7110-94) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET7110-94_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-VPS28 antibody (ET7110-94) at 1/200 dilution.

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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-94) at 1/200 dilution for 0.5 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.
ET7110-94_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue with Rabbit anti-VPS28 antibody (ET7110-94) at 1/200 dilution.

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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-94) at 1/200 dilution for 0.5 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.
ET7110-94_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human uterus tissue with Rabbit anti-VPS28 antibody (ET7110-94) at 1/200 dilution.

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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-94) at 1/200 dilution for 0.5 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.
ET7110-94_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-VPS28 antibody (ET7110-94) at 1/50 dilution.

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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-94) at 1/50 dilution for 0.5 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.
ET7110-94_7.jpg Fig7: Flow cytometric analysis of VPS28 was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7110-94, 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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