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, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | JE55-08 |
| 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*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: 25 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within Human VPS28 aa 100-221 / 221. |
| Positive control: | HepG2 cell lysate, Jurkat cell lysate, A549 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, Jurkat. |
| Subcellular location: | Late endosome membrane, Cell membrane. |
| Recommended Dilutions:
WB IF-Cell FC IP |
1:500-1:2,000 1:50-1:200 1:1,000 1-2μg/sample |
| 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
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Fig1:
Western blot analysis of VPS28 on different lysates with Rabbit anti-VPS28 antibody (ET7110-94) at 1/2,000 dilution. Lane 1: HepG2 cell lysate Lane 2: Jurkat cell lysate Lane 3: A549 cell lysate Lane 4: Mouse brain tissue lysate Lane 5: Rat brain tissue lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 25 kDa Observed band size: 25 kDa Exposure time: 3 minutes; 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 (ET7110-94) 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. |
|
Fig2:
Immunocytochemistry analysis of Jurkat cells labeling VPS28 with Rabbit anti-VPS28 antibody (ET7110-94) 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-VPS28 antibody (ET7110-94) 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:
Flow cytometric analysis of Jurkat cells labeling VPS28. Cells were fixed and permeabilized. Then stained with the primary antibody (ET7110-94, 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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Fig4:
VPS28 was immunoprecipitated from 0.2 mg Jurkat cell lysate with ET7110-94 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET7110-94 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: Jurkat cell lysate (input) Lane 2: ET7110-94 IP in Jurkat cell lysate Lane 3: Rabbit IgG instead of ET7110-94 in Jurkat cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 12 seconds; ECL: K1801 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.