Genetics of Parkinson's Disease: Lysosomal Dysfunction Antibody Sampler Kit
cat.: HAK21127
| Product Type: | Antibody Sampler Kit |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P |
| 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. |
| Uniprot #: | SwissProt: P04062 Human | P17439 Mouse | P07339 Human | P18242 Mouse | P24268 Rat | Q9NQ11 Human | Q9CTG6 Mouse | Q96QK1 Human | Q9EQH3 Mouse | G3V8A5 Rat | I0FFY6 Monkey | Q5S007 Human | Q5S006 Mouse | P37840 Human | O55042 Mouse | P37377 Rat Entrez Gene: 684536 Rat | 362645 Rat | 300160 Rat |
Images
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Fig1:
Western blot analysis of VPS35 on different lysates with Rabbit anti-VPS35 antibody (HA723858) at 1/5,000 dilution. Lane 1: SH-SY5Y cell lysate (20 µg/Lane) Lane 2: A549 cell lysate (20 µg/Lane) Lane 3: U-937 cell lysate (20 µg/Lane) Lane 4: COS-1 cell lysate (20 µg/Lane) Lane 5: Neuro-2a cell lysate (20 µg/Lane) Lane 6: NIH/3T3 cell lysate (20 µg/Lane) Lane 7: C2C12 cell lysate (20 µg/Lane) Lane 8: C6 cell lysate (20 µg/Lane) Lane 9: NR8383 cell lysate (20 µg/Lane) Lane 10: PC-12 cell lysate (20 µg/Lane) Lane 11: Mouse brain tissue lysate (40 µg/Lane) Lane 12: Mouse colon tissue lysate (40 µg/Lane) Lane 13: Rat brain tissue lysate (40 µg/Lane) Lane 14: Rat colon tissue lysate (40 µg/Lane) Predicted band size: 92 kDa Observed band size: 80 kDa Exposure time: 59 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 (HA723858) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Western blot analysis of VPS35 on different lysates with Rabbit anti-VPS35 antibody (ET7107-01) at 1/1,000 dilution. Lane 1: U-937 cell lysate (20 µg/Lane) Lane 2: A549 cell lysate (20 µg/Lane) Lane 3: NIH/3T3 cell lysate (20 µg/Lane) Lane 4: C2C12 cell lysate (20 µg/Lane) Lane 5: Mouse colon tissue lysate (40 µg/Lane) Predicted band size: 92 kDa Observed band size: 80 kDa Exposure time: 30 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 (ET7107-01) at 1/1,000 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. |
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Fig3:
Immunocytochemistry analysis of Neuro-2a cells labeling VPS35 with Rabbit anti-VPS35 antibody (HA723858) 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 (HA723858) 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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Fig4:
Western blot analysis of VPS35 on different lysates with Rabbit anti-VPS35 antibody (ET7107-01) at 1/2,000 dilution. Lane 1: HAP1-parental cell lysate Lane 2: HAP1-VPS35 KD cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 92 kDa Observed band size: 80 kDa Exposure time: 20 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 (ET7107-01) at 1/2,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig5:
Western blot analysis of ATP13A2 on different lysates with Rabbit anti-ATP13A2 antibody (HA722741) at 1/1,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: SH-SY5Y cell lysate (20 µg/Lane) Lane 3: Neuro-2a cell lysate (20 µg/Lane) Lane 4: C6 cell lysate (20 µg/Lane) Lane 5: COS-1 cell lysate (20 µg/Lane) Predicted band size: 129 kDa Observed band size: 160 kDa Exposure time: 40 seconds; ECL: K1802; 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 (HA722741) at 1/1,000 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. |
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Fig6:
Western blot analysis of Cathepsin D on different lysates with Rabbit anti-Cathepsin D antibody (ET1608-49) at 1/2,000 dilution. Lane 1: HepG2 cell lysate (15 µg/Lane) Lane 2: MCF7 cell lysate (15 µg/Lane) Lane 3: Neuro-2a cell lysate (15 µg/Lane) Lane 4: Mouse brain tissue lysate (20 µg/Lane) Predicted band size: 45/28 kDa Observed band size: 45/28 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 (ET1608-49) 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. |
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Fig7:
Western blot analysis of Cathepsin D on different lysates with Rabbit anti-Cathepsin D antibody (ET1608-49) at 1/1,000 dilution. Lane 1: C6 cell lysate (20 µg/Lane) Lane 2: Mouse brain tissue lysate (20 µg/Lane) Lane 3: Rat brain tissue lysate (20 µg/Lane) Predicted band size: 45/28 kDa Observed band size: 45/28 kDa Exposure time: 6 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 (ET1608-49) at 1/1,000 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. |
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Fig8:
Immunofluorescence analysis of frozen mouse hippocampus tissue with Rabbit anti-ATP13A2 antibody (HA722741) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA722741, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
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Fig9:
Immunofluorescence analysis of frozen mouse hippocampus tissue with Rabbit anti-ATP13A2 antibody (HA722741) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA722741, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
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Fig10:
Immunofluorescence analysis of frozen mouse cerebral cortex tissue with Rabbit anti-ATP13A2 antibody (HA722741) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA722741, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
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Fig11:
Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-ATP13A2 antibody (HA722741) 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 (HA722741) 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. |
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Fig12:
Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-LRRK2 antibody (HA722932) 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 (HA722932) 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. |
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Fig13:
Immunofluorescence analysis of frozen mouse cerebellum tissue with Rabbit anti-Alpha-Synuclein antibody (HA723035) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA723035, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
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Fig14:
Immunofluorescence analysis of frozen rat cerebellum tissue with Rabbit anti-Alpha-Synuclein antibody (HA723035) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA723035, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
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Fig15:
Western blot analysis of VPS35 on different lysates with Rabbit anti-VPS35 antibody (HA723858) at 1/5,000 dilution. Lane 1: HAP1-parental cell lysate (10 µg/Lane) Lane 2: HAP1-VPS35 KD cell lysate (10 µg/Lane) Predicted band size: 92 kDa Observed band size: 80 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 (HA723858) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig16:
VPS35 was immunoprecipitated from 0.2 mg A549 cell lysate with HA723858 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723858 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: HA723858 IP in A549 cell lysate Lane 3: Rabbit IgG instead of HA723858 in A549 cell lysate Blocking/Dilution buffer: primary antibody dilution (K1803) Exposure time: 3 minutes; ECL: K1801 |
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