MA5-45942

antibody from Invitrogen Antibodies

Targeting: VPS35 FLJ10752, MEM3, PARK17

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-45942

Provider

Invitrogen Antibodies

Product name

VPS35 Monoclonal Antibody (8A3), APC

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Reactivity

Human, Mouse, Rat

Host

Mouse

Isotype

IgG

Antibody clone number

8A3

Vial size

100 µg

Concentration

1 mg/mL

Storage

4° C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of Human SH-SY5Y cells showing detection of VPS35 protein. Lane 1: Molecular Weight Ladder. Lane 2: SH-SY5Y cells (10 µg). Load: 10 µg. Blocking: 5% Skim Milk powder in TBST. Samples were incubated with VPS35 monoclonal antibody (Product # MA5-45942) at 1:1,000 for 2 hours at RT with shaking, followed by Goat anti-mouse IgG:HRP at 1:4,000 for 1 hour at RT with shaking. Color Development: Chemiluminescent for HRP (Moss) for 5 min in RT.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of Human, Mouse A549, MEF showing detection of VPS35 protein. Lane 1: Molecular Weight Ladder. Lane 2: VPS35 KO A549 cells. Lane 3: mouse embryonic fibroblast cells. Load: 8 µg each A549 and MEF. Samples were incubated with VPS35 monoclonal antibody (Product # MA5-45942) at 1:5 (tissue culture supernatant), followed by Donkey anti-mouse IRDye 800CW at 1:25,000 in TBS-T.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry/Immunofluorescence analysis using A549 WT, VPS35 KO cells. Samples were incubated with VPS35 monoclonal antibody (Product # MA5-45942), followed by Donkey Anti-Mouse AlexaFluor 594.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis using Kidney. Samples were incubated with VPS35 monoclonal antibody (Product # MA5-45942) at 1:100 for Overnight at 4C, then 30 min at 37C, followed by Goat Anti-Mouse IgG (H+L): FITC for 45 min at 37C. Counterstain used was DAPI for 3 min at RT. Magnification: 20X.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoprecipitation analysis using mouse embryonic fibroblast. Samples were incubated with VPS35 monoclonal antibody (Product # MA5-45942). Three amounts of (Product # MA5-45942) (3, 1 and 0.3 µg) were non-covalently coupled to 10 µL of A/G sepharose beads for 1 hour at 4°C and next incubated with 250 µg of MEF lysate for 2 hours at 4°C.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoprecipitation analysis using A549 cells. 500 µL cell culture supernatants were incubated with 10 µL of Protein A/G resin beads for 1 hour at 4⁰C. VPS35 monoclonal antibody (Product # MA5-45942) depletes VPS35 from the A549 cell extract.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoprecipitation analysis using A549 cells. Three amounts of (Product # MA5-45942) (3, 1 and 0.3 µg) were non-covalently coupled to 10 µL of A/G sepharose beads for 1 hour at 4°C and next incubated with 250 µg of A549 lysate for 2 hours at 4°C.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoprecipitation analysis using A549 cells. Samples incubated with VPS35 monoclonal antibody (Product # MA5-45942).