VPS34 Antibody

Novus Biologicals | Catalog # NB110-87320

Novus Biologicals
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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western, Knockdown Validated

Cited:

Western Blot, Flow Cytometry

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG
View all VPS34/PIK3C3 Primary Antibodies »

Product Specifications

Immunogen

Synthetic peptide made to an internal portion of the human VPS34 protein (within residues 700-850). [Swiss-Prot# Q8NEB9]

Reactivity Notes

Immunogen displays the following percentage of sequence identity for non-tested species: Xenopus (87%).

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

100 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for VPS34 Antibody

Simple Western: VPS34 Antibody [NB110-87320]

Simple Western: VPS34 Antibody [NB110-87320]

Simple Western: VPS34 Antibody [NB110-87320] - Simple Western lane view shows a specific band for VPS34 in 0.5 mg/ml of HepG2 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.
Knockdown Validated: VPS34 Antibody [NB110-87320]

Western Blot: VPS34 Antibody [NB110-87320]

VPS34-Antibody-Knockdown-Validated-NB110-87320-img0007.jpg
Western Blot: VPS34 Antibody [NB110-87320]

Western Blot: VPS34 Antibody [NB110-87320]

Western Blot: VPS34 Antibody [NB110-87320] - HepG2 whole cell lysates.
Immunocytochemistry/ Immunofluorescence: VPS34 Antibody [NB110-87320]

Immunocytochemistry/ Immunofluorescence: VPS34 Antibody [NB110-87320]

Immunocytochemistry/Immunofluorescence: VPS34 Antibody [NB110-87320] - VPS34 antibody was tested at 1:200 in HeLa cells with FITC (green). Nuclei (Blue) were counterstained with Dapi (blue).
VPS34 Antibody

Western Blot: VPS34 Antibody [NB110-87320] -

Western Blot: VPS34 Antibody [NB110-87320] - Celastrol regulates autophagy- & mitophagy-related gene expressions in neurons. SH-SY5Y cells treated with 0.1–1 μM celastrol for 4 h. (A) Real-time quantitative PCR results show that 1 μM celastrol treatment for 4 h enhanced mRNA expressions of phagophore induction genes Beclin 1 & Ambra1, phagophore elongation genes Atg7 & Atg12, & vacuole formation genes LC3A & Atg4B. (B) Western blot results of SH-SY5Y treated with 100–500 nM celastrol show celastrol (500 nM) increased in protein expressions of Beclin 1 & Vps34 (phagophore induction), Atg7 (phagophore elongation), LC3-II (vacuole formation), DJ-1, & PINK1 (mitophagy). Celastrol (500 nM) suppressed LRRK2 expression. (C) MPTP (10 mg/kg/day for 3 days, i.p.) but not celastrol (3 mg/kg/day for 3 days, i.p.) caused dopaminergic nerve terminal degeneration (tyrosine hydroxylase↓) & mitophagy inactivation (DJ-1↓ & PINK1↓) in the striatum of mice as compared to saline control. Celastrol cotreatment with MPTP reversed it as compared to MPTP. Data are given as the mean ± SEM (n = 3–5). p-value was determined using the Kruskal–Wallis test followed by Dunn’s multiple comparison post hoc test. * p < 0.05 compared with controls. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31906147), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for VPS34 Antibody

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:100

Simple Western

1:20

Western Blot

1:1000
Application Notes

This VPS34 antibody is useful for Immunocytochemistry/Immunofluorescence and Western blot, where a band is seen approx. 100 kDa.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in HepG2 lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:20, apparent MW was 102 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reviewed Applications

Read 1 review rated 4 using NB110-87320 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS, 0.1% BSA, and 50% Glycerol

Preservative

0.05% Sodium Azide

Concentration

0.2 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: VPS34/PIK3C3

VPS34 (vacuolar protein sorting 34) is a member of PI3K family of lipid kinases, all of which phosphorylate the 3' hydroxy position of phosphatidylinositol ring. VPS34 is ubiquitously expressed with highest expression in skeletal muscles and it localizes to midbody/late endosomes in cells. VPS34 forms a complexes with BECN1, PIK3R4 and either UVRAG and KIAA0226/Rubicon, or with ATG14, and the presence of UVRAG/ATG14 are mutually exclusive in this complex. It forms complex with PIK3R4/PIK3CB also, and interacts with RAB7A in the presence of PIK3R4. VPS34 regulates vesicular trafficking in the endosomal/lysosomal system, where it is involved in the recruitment of proteins containing phosphatidylinositol 3-phosphate (PtdIns3P) binding domains to intracellular membranes. As PI3K complex's catalytic subunit, VPS34 mediates formation of PtdIns3P which is involved in initiation and maturation of autophagosomes. Moreover, VPS34 is implicated in transporting lysosomal enzyme precursors to lysosomes, required for abcission step in cytokinesis and transport from early to late endosomes. Vps34 has also been involved in other signalling processes such as nutrient sensing in mTOR pathway in mammalian cells and trimeric G-protein signalling to MAPK in yeast.

Alternate Names

PI 3-Kinase C3, PI3K type 3, PIK3C3

Entrez Gene IDs

5289 (Human); 225326 (Mouse); 65052 (Rat)

Gene Symbol

PIK3C3

UniProt

Q8NEB9 (Human)

Additional VPS34/PIK3C3 Products

Product Documents for VPS34 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for VPS34 Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Customer Reviews for VPS34 Antibody (1)

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  • VPS34 Antibody
    Name: Naoya Nakai
    Application: Western Blot
    Sample Tested: Skeletal muscle satellite cells (cultured in vitro)
    Species: Mouse
    Verified Customer | Posted 07/27/2017
    Lane 1 - 4 : Cytoplasmic fraction Lane 5 - 8 : Membrane fraction
    VPS34 Antibody NB110-87320

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Protocols

View specific protocols for VPS34 Antibody (NB110-87320):

VPS34 Antibody:
Western Blot Protocol

1. Perform SDS-PAGE (4-12% MOPS) on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer
apparatus.
3. Rinse membrane with dH2O and then stain the blot using Ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
4. Rinse the blot in TBS for approximately 5 minutes.
5. Block the membrane using 5% BSA in TBS + Tween, 1 hour at RT.
6. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
7. Dilute the rabbit anti-VPS34 primary antibody (NB 110-87320) in blocking buffer and incubate 1 hour at room temperature.
8. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
9. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturers
instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions (Pierce ECL).

Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%, provided it does not interfere with antibody-antigen binding.

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