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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
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- Product number
- PA1-30301 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- VPS34 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Recommended positive controls: HeLa cell lysate.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 400 µL
- Concentration
- 1.4 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of PI 3 Kinase 3 in HeLa cell lysate using a PI 3 Kinase 3 polyclonal antibody (Product # PA1-30301) followed by detection using an HRP-anti-Rabbit secondary antibody and chemiluminescence.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of VPS34 was performed by loading 20 µg per lane of HeLa, SH-SY5Y cell lysates. Proteins were transferred to a membrane and probed with a VPS34 Polyclonal Antibody (Product # PA1-30301) at a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-VPS34 Polyclonal Antibody (Product # PA1-30301) and a 102 kDa band corresponding to Phosphatidylinositol 3-kinase catalytic subunit type 3 was observed across cell lines. Whole cell extracts (30 µg lysate) of Reh (Lane 1), A-375 (Lane 2), HT-1080 (Lane 3), U-2 OS (Lane 4), HT-29 (Lane 5), PC-3 (Lane 6), Hep G2 (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1,000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry-Immunofluorescence analysis of VPS34 in HeLa cells using VPS34 Polyclonal Antibody (Product # PA1-30301) (Green). Blue: DAPI.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Phosphatidylinositol 3-kinase catalytic subunit type 3 was performed using 70% confluent log phase A-375 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with VPS34 Polyclonal Antibody (Product # PA1-30301) at 1:50 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Membrane localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry (Paraffin) analysis of VPS34 in human hepatocarcinoma tissue using VPS34 Polyclonal Antibody (Product # PA1-30301).
