Product nameAnti-VPS35 antibody
See all VPS35 primary antibodies
DescriptionRabbit polyclonal to VPS35
Tested applicationsSuitable for: WB, ICC/IF, IHC-Pmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat, Rabbit, Horse, Cow, Dog, Pig, Chimpanzee, Macaque monkey, Orangutan
Synthetic peptide conjugated to KLH derived from within residues 400 - 500 of Human VPS35.
- This antibody gave a positive signal in the following whole cell lysates: HepG2; MOLT4; Raji. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal heart muscle. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: HepG2.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab118838 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 91 kDa (predicted molecular weight: 91 kDa).|
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-P||Use a concentration of 5 µg/ml.|
FunctionEssential component of the retromer complex, a complex required to retrieve lysosomal enzyme receptors (IGF2R and M6PR) from endosomes to the trans-Golgi network. Also required to regulate transcytosis of the polymeric immunoglobulin receptor (pIgR-pIgA).
Tissue specificityUbiquitous. Highly expressed in heart, brain, placenta, skeletal muscle, spleen, thymus, testis, ovary, small intestine, kidney and colon.
Sequence similaritiesBelongs to the VPS35 family.
Cellular localizationCytoplasm. Membrane.
- Information by UniProt
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: VPS35 knockout HAP1 cell lysate (20 µg)
Lane 3: NIH/3T3 cell lysate (20 µg)
Lane 4: A549 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab118838 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab118838 was shown to recognize VPS35 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when VPS35 knockout samples were examined. Wild-type and VPS35 knockout samples were subjected to SDS-PAGE. ab118838 and ab8245 (loading control to GAPDH) were diluted 1μg/mL and 1/2000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
ICC/IF image of ab118838 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab118838 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
All lanes : Anti-VPS35 antibody (ab118838) at 1 µg/ml
Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 2 : MOLT4 (Human acute lymphoblastic leukemia cell line) Whole Cell Lysate
Lane 3 : Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 91 kDa
Observed band size: 85 kDa why is the actual band size different from the predicted?
Additional bands at: 24 kDa, 35 kDa, 71 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 2 minutes
ab118838 has not yet been referenced specifically in any publications.