VPS35 Recombinant Rabbit Monoclonal Antibody [JB33-82]
cat.: ET7107-01
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Cynomolgus monkey, Pig
Applications: WB, IHC-P, IF-Cell
Clonality: Monoclonal
Clone number: JB33-82
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: 92 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human VPS35 aa 80-220 / 796.
Positive control: U-937 cell lysate, A549 cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, mouse colon tissue lysate, L6 cell lysate, rat colon tissue lysate, human colon cancer tissue, human kidney tissue, human placenta tissue, mouse colon tissue, NIH/3T3.
Subcellular location: Cytoplasm. Membrane. Endosome.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
1:2,000-1:5,000
1:100-1:500
1:100
Uniprot #: SwissProt: Q96QK1 Human | Q9EQH3 Mouse
Entrez Gene: 25479 Rat
Alternative names: DKFZp434E1211 DKFZp434P1672 FLJ10752 FLJ13588 FLJ20388 hVPS35 Maternal embryonic 3 Maternal-embryonic 3 MEM 3 MEM3 PARK17 TCCCTA00141 Vacuolar protein sorting 35 (yeast) Vacuolar protein sorting 35 Vacuolar protein sorting 35 homolog Vacuolar protein sorting associated protein 35 Vacuolar protein sorting-associated protein 35 Vesicle protein sorting 35 VPS 35 VPS35 VPS35_HUMAN
Images
ET7107-01_1.jpg Fig1: Western blot analysis of VPS35 on different lysates with Rabbit anti-VPS35 antibody (ET7107-01) at 1/2,000 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 1/2,000 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.
ET7107-01_2.jpg Fig2: 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.
ET7107-01_3.jpg Fig3: Western blot analysis of VPS35 on different lysates with Rabbit anti-VPS35 antibody (ET7107-01) at 1/2,000 dilution.

Lane 1: L6 cell lysate (10 µg/Lane)
Lane 2: Rat colon tissue lysate (20 µg/Lane)

Predicted band size: 92 kDa
Observed band size: 80 kDa

Exposure time: 24 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 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.
ET7107-01_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-VPS35 antibody. Counter stained with hematoxylin.
ET7107-01_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-VPS35 antibody. Counter stained with hematoxylin.
ET7107-01_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-VPS35 antibody. Counter stained with hematoxylin.
ET7107-01_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-VPS35 antibody. Counter stained with hematoxylin.
ET7107-01_8.jpg Fig8: Immunocytochemistry analysis of NIH/3T3 cells labeling VPS35 with Rabbit anti-VPS35 antibody (ET7107-01) 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 (ET7107-01) 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.