WB
Western blot analysis of VPS28 on different lysates with Rabbit anti-VPS28 antibody 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. Exposure time: 3 minutes; 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.ICC/IF
Immunocytochemistry analysis of Jurkat cells labeling VPS28 with Rabbit anti-VPS28 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-VPS28 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 Jurkat cells labeling VPS28. 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
VPS28 was immunoprecipitated from 0.2 mg Jurkat cell lysate with Rabbit anti-VPS28 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-VPS28 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: Jurkat cell lysate (input), Lane 2: Rabbit anti-VPS28 antibody IP in Jurkat cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-VPS28 antibody in Jurkat cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 12 seconds.| Product Name | VPS28 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within Human VPS28 aa 100-221 / 221. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIPWB |
| WB:1:500-1:2000 ICC/IF:1:50-1:200 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | VPS28 |
|---|---|
| Gene Synonyms | CIIA |
| Gene Full Name | VPS28 subunit of ESCRT-I |
| Gene Summary | This gene encodes a protein subunit of the ESCRT-I complex (endosomal complexes required for transport), which functions in the transport and sorting of proteins into subcellular vesicles. This complex can also be hijacked to facilitate the budding of enveloped viruses from the cell membrane. Alternative splicing results in multiple transcript variants encoding different isoforms. [provided by RefSeq, Jul 2013] |
| Alternative Names | ESCRT-I complex subunit VPS28 antibody H-Vps28 antibody Vacuolar protein sorting 28 antibody Vacuolar protein sorting 28 (yeast) antibody Vacuolar protein sorting 28 homolog (S. cerevisiae) antibody Vacuolar protein sorting 28 homolog antibody Vacuolar protein sorting-associated protein 28 homolog antibody VPS 28 antibody Vps28 antibody VPS28_HUMAN antibody ESCRT-I complex subunit VPS28 antibody H-Vps28 antibody Vacuolar protein sorting 28 antibody Vacuolar protein sorting 28 (yeast) antibody Vacuolar protein sorting 28 homolog (S. cerevisiae) antibody Vacuolar protein sorting 28 homolog antibody Vacuolar protein sorting-associated protein 28 homolog antibody VPS 28 antibody Vps28 antibody VPS28_HUMAN antibody Yeast class E protein Vps28p homolog antibody |
| Molecular Weight(MW) | 25kDa |
| Cellular Localization | Late endosome membrane, Cell membrane. |
WB
Western blot analysis of VPS28 on different lysates with Rabbit anti-VPS28 antibody 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. Exposure time: 3 minutes; 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.
ICC/IF
Immunocytochemistry analysis of Jurkat cells labeling VPS28 with Rabbit anti-VPS28 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-VPS28 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 Jurkat cells labeling VPS28. 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
VPS28 was immunoprecipitated from 0.2 mg Jurkat cell lysate with Rabbit anti-VPS28 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-VPS28 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: Jurkat cell lysate (input), Lane 2: Rabbit anti-VPS28 antibody IP in Jurkat cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-VPS28 antibody in Jurkat cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 12 seconds.| Application Notes | WB:1:500-1:2000 ICC/IF:1:50-1:200 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | 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. |
