WB
Western blot analysis of VPS35 on different lysates with Rabbit anti-VPS35 antibody at 1/5,000 dilution. Exposure time: 59 seconds; 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 1/5,000 dilution was used in primary antibody dilution 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 SH-SY5Y cells labeling VPS35 with Rabbit anti-VPS35 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-VPS35 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.ICC/IF
Immunocytochemistry analysis of Neuro-2a cells labeling VPS35 with Rabbit anti-VPS35 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-VPS35 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.ICC/IF
Immunocytochemistry analysis of C6 cells labeling VPS35 with Rabbit anti-VPS35 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-VPS35 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.IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-VPS35 antibody at 1/200 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 at 1/200 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.IHC
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-VPS35 antibody at 1/200 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 at 1/200 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.IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-VPS35 antibody at 1/200 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 at 1/200 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.FC
Flow cytometric analysis of SH-SY5Y cells labeling VPS35. 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
VPS35 was immunoprecipitated from 0.2 mg A549 cell lysate with VPS35 Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using VPS35 Recombinant Rabbit Monoclonal Antibody at 1/5,000 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: A549 cell lysate (input) Lane 2: VPS35 Recombinant Rabbit Monoclonal Antibody IP in A549 cell lysate Lane 3: Rabbit IgG instead of VPS35 Recombinant Rabbit Monoclonal Antibody in A549 cell lysate Blocking/Dilution buffer: primary antibody dilution, Exposure time: 3 minutes;IP
VPS35 was immunoprecipitated from 0.2 mg NIH/3T3 cell lysate with VPS35 Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using VPS35 Recombinant Rabbit Monoclonal Antibody at 1/5,000 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: NIH/3T3 cell lysate (input) Lane 2: VPS35 Recombinant Rabbit Monoclonal Antibody IP in NIH/3T3 cell lysate Lane 3: Rabbit IgG instead of VPS35 Recombinant Rabbit Monoclonal Antibody in NIH/3T3 cell lysate Blocking/Dilution buffer: primary antibody dilution. Exposure time: 46 seconds.| Product Name | VPS35 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within human VSP35 aa 760-795. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:5000 ICC/IF:1:100 IHC:1:200-1:1000 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMonkeyMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | VPS35 |
|---|---|
| Gene Synonyms | MEM3 PARK17 |
| Gene Full Name | VPS35 retromer complex component |
| Gene Summary | This gene belongs to a group of vacuolar protein sorting (VPS) genes. The encoded protein is a component of a large multimeric complex, termed the retromer complex, involved in retrograde transport of proteins from endosomes to the trans-Golgi network. The close structural similarity between the yeast and human proteins that make up this complex suggests a similarity in function. Expression studies in yeast and mammalian cells indicate that this protein interacts directly with VPS35, which serves as the core of the retromer complex. [provided by RefSeq, Jul 2008] |
| Alternative Names | DKFZp434E1211 antibody DKFZp434P1672 antibody FLJ10752 antibody FLJ13588 antibody FLJ20388 antibody hVPS35 antibody Maternal embryonic 3 antibody Maternal-embryonic 3 antibody MEM 3 antibody MEM3 antibody PARK17 antibody TCCCTA00141 antibody Vacuolar protein sorting 35 antibody Vacuolar protein sorting 35 homolog antibody Vacuolar protein sorting associated protein 35 antibody Vacuolar protein sorting-associated protein 35 antibody Vesicle protein sorting 35 antibody VPS 35 antibody |
| Molecular Weight(MW) | 92 kDa(Observed band size: 80 kDa) |
| Cellular Localization | Cytoplasm, Membrane, Endosome, Early endosome, Late endosome. |
WB
Western blot analysis of VPS35 on different lysates with Rabbit anti-VPS35 antibody at 1/5,000 dilution. Exposure time: 59 seconds; 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 1/5,000 dilution was used in primary antibody dilution 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 SH-SY5Y cells labeling VPS35 with Rabbit anti-VPS35 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-VPS35 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.
ICC/IF
Immunocytochemistry analysis of Neuro-2a cells labeling VPS35 with Rabbit anti-VPS35 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-VPS35 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.
ICC/IF
Immunocytochemistry analysis of C6 cells labeling VPS35 with Rabbit anti-VPS35 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-VPS35 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.
IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-VPS35 antibody at 1/200 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 at 1/200 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.
IHC
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-VPS35 antibody at 1/200 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 at 1/200 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-VPS35 antibody at 1/200 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 at 1/200 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.
FC
Flow cytometric analysis of SH-SY5Y cells labeling VPS35. 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
VPS35 was immunoprecipitated from 0.2 mg A549 cell lysate with VPS35 Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using VPS35 Recombinant Rabbit Monoclonal Antibody at 1/5,000 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: A549 cell lysate (input) Lane 2: VPS35 Recombinant Rabbit Monoclonal Antibody IP in A549 cell lysate Lane 3: Rabbit IgG instead of VPS35 Recombinant Rabbit Monoclonal Antibody in A549 cell lysate Blocking/Dilution buffer: primary antibody dilution, Exposure time: 3 minutes;
IP
VPS35 was immunoprecipitated from 0.2 mg NIH/3T3 cell lysate with VPS35 Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using VPS35 Recombinant Rabbit Monoclonal Antibody at 1/5,000 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: NIH/3T3 cell lysate (input) Lane 2: VPS35 Recombinant Rabbit Monoclonal Antibody IP in NIH/3T3 cell lysate Lane 3: Rabbit IgG instead of VPS35 Recombinant Rabbit Monoclonal Antibody in NIH/3T3 cell lysate Blocking/Dilution buffer: primary antibody dilution. Exposure time: 46 seconds.| Application Notes | WB:1:5000 ICC/IF:1:100 IHC:1:200-1:1000 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | 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 | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
