VPS35 Antibody: RPE

SKU: orb612806

Description

Mouse monoclonal antibody against VPS35 conjugated to RPE

Images & Validation

• Tested Applications: ICC, IF, IHC, WB
• Dilution range: WB (1:1000); ICC/IF (1:200); IP (1:200); IHC (1:100)
• Reactivity: Human, Mouse, Rat
• Application Notes:
  A 1:1000 dilution of SMC-604 was sufficient for detection of VPS35 in 10 µg of SH-SY5Y by ECL immunoblot analysis using Goat Anti-Mouse IgG:HRP as the secondary antibody.

Key Properties

• Host: Mouse
• Clonality: Monoclonal
• Isotype: IgG1
• Clone No.: 8A3
• Immunogen: Full length recombinant human VSP35
• Target: VPS35
• Molecular Weight: 92 kDa
• Purification: Protein G Purified

Storage & Handling

• Storage: Conjugated antibodies should be stored according to the product label
• Buffer/Preservatives: 95.64mM Phosphate, 2.48mM MES and 2mM EDTA
• Concentration: 1 mg/ml
• Disclaimer: For research use only

Alternative Names

Vacuolar protein sorting-associated protein 35, MEM3, PARK17, VPS35 retromer complex component, maternal-embryonic 3, vesicle protein sortin 35, TCCCTA00141, FLJ10752

Immunohistochemistry analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 8A3. Tissue: Kidney. Species: Mouse. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody at 1:100 for Overnight at 4°C, then 30 min at 37°C. Secondary Antibody: Goat Anti-Mouse IgG (H+L): FITC for 45 min at 37°C. Counterstain: DAPI for 3 min at RT. Magnification: 20X.

Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 8A3. Tissue: A549 cells. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody at 1:5 (tissue culture supernatant). Secondary Antibody: Donkey anti-mouse: Alexa Fluor 594 at 1:4000 in 0.2% BSA PBS. Counterstain: DAPI. Localization: Vesicles. A) VPS35 KO A549 cells B) WT A549 cells.

Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 8A3. Tissue: A549 WT, VPS35 KO cells. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody. Secondary Antibody: Donkey Anti-Mouse AlexaFluor 594. Clone can detect VPS35 at 1/2000 concentration.

Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 8A3. Tissue: A549 cells. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody.

Western Blot analysis of Human, Mouse A549, MEF showing detection of VPS35 protein using Mouse Anti-VPS35 Monoclonal Antibody, Clone 8A3. Lane 1: Molecular Weight Ladder. Lane 2: VPS35 KO A549 cells. Lane 3: mouse embryonic fibroblast cells. Load: 8 μg each A549 and MEF. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody at 1:5 (tissue culture supernatant). Secondary Antibody: Donkey anti-mouse IRDye 800CW at 1:25000 in TBS-T.

Western Blot analysis of Human SH-SY5Y showing detection of VPS35 protein using Mouse Anti-VPS35 Monoclonal Antibody, Clone 8A3. Lane 1: Molecular Weight Ladder. Lane 2: SH-SY5Y (10 ug). Load: 10 μg. Block: 5% Skim Milk powder in TBST. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody at 1:1000 for 2 hours at RT with shaking. Secondary Antibody: Goat anti-mouse IgG:HRP at 1:4000 for 1 hour at RT with shaking. Color Development: Chemiluminescent for HRP (Moss) for 5 min in RT.

Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 8A3. Tissue: A549 cells. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody. Three amounts of (3, 1 and 0.3 ug) were non-covalently coupled to 10uL of A/G sepharose beads for 1 hour at 4°C and next incubated with 250ug of A549 lysate for 2 hours at 4°C.

Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 8A3. Tissue: embryonic fibroblast. Species: Mouse. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody. Three amounts of (3, 1 and 0.3 ug) were non-covalently coupled to 10uL of A/G sepharose beads for 1 hour at 4°C and next incubated with 250ug of MEF lysate for 2 hours at 4°C.

Immunoprecipitation analysis using Mouse Anti-VPS35 Monoclonal Antibody, Clone 8A3. Tissue: A549 cells. Species: Human. Primary Antibody: Mouse Anti-VPS35 Monoclonal Antibody. 500 μL cell culture supernatants were incubated with 10 μL of Protein A/G resin beads for 1 hour at 4°C.

Quick Database Links

Gene Symbol

VPS35

UniProt

Q96QK1

NCBI Gene ID

55737

RefSeq (Protein):NP_060676.2