Anti-VPS35 antibody [EPR11501(B)] (ab157220)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR11501(B)] to VPS35
- Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
Overview
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Product name
Anti-VPS35 antibody [EPR11501(B)]
See all VPS35 primary antibodies -
Description
Rabbit monoclonal [EPR11501(B)] to VPS35 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF HumanIHC-P HumanWB Human -
Immunogen
Synthetic peptide within Human VPS35. The exact sequence is proprietary.
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Positive control
- WB: HeLa, HepG2, Raji and MOLT4 cell lysates. IHC-P: Human kidney and human ovarian carcinoma tissue. ICC/IF: MOLT4 and wild-type HAP1 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EPR11501(B) -
Isotype
IgG -
Research areas
Images
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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 - ab157220 observed at 91 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab157220 was shown to specifically react with VPS35 in wild-type HAP1 cells. No band was observed when VPS35 knockout samples were examined. Wild-type and VPS35 knockout samples were subjected to SDS-PAGE. ab157220 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively 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. -
ab157220 staining VPS35 in wild-type HAP1 cells (top panel) and VPS35 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab157220 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
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All lanes : Anti-VPS35 antibody [EPR11501(B)] (ab157220) at 1/10000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : HepG2 cell lysate
Lane 3 : Raji cell lysate
Lane 4 : MOLT4 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-rabbit HRP at 1/2000 dilution
Predicted band size: 92 kDa
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Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling VPS35 with ab157220 at 1/50 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human ovarian carcinoma tissue labeling VPS35 with ab157220 at 1/50 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Flow cytometric analysis of permeabilized MOLT4 cells labeling VPS35 with ab157220 at 1/10 dilution (red) or a rabbit IgG (negative) (green).
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