PA5-80208
antibody from Invitrogen Antibodies
Targeting: UPF1
HUPF1, KIAA0221, NORF1, pNORF1, RENT1, smg-2
Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [5]
- Immunocytochemistry [1]
- Immunohistochemistry [3]
- Flow cytometry [1]
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Validation data
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- Product number
- PA5-80208 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- UPF1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Reconstitute with 0.2 mL of distilled water to yield a concentration of 500 µg/mL.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 500 µg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of UPF1 in Lane 1: rat Pancreas tissue lysate, Lane 2: PANC whole cell lysate using 40-50 µg per well. Sample was incubated with UPF1 (Product # PA5-80208) at a dilution of 0.5 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of UPF1 in Lane 1: rat Pancreas tissue lysate, Lane 2: PANC whole cell lysate using 40-50 µg per well. Sample was incubated with UPF1 (Product # PA5-80208) at a dilution of 0.5 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of RENT1/hUPF1 in, Lane 1: human Hela whole cell lysates, Lane 2: human Raji whole cell lysates, Lane 3: human HepG2 whole cell lysates, Lane 4: human SK-OV-3 whole cell lysates, Lane 5: human PC-3 whole cell lysates, Lane 6: human HEK293 whole cell lysates, Lane 7: rat RH35 whole cell lysates, Lane 8: mouse HEPA1-6 whole cell lysates. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 µg of sample under reducing conditions. After Electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. The membrane was blocked with 5% non-fat milk/TBS for 1. 5 hour at RT. The membrane was incubated with UPF1 Polyclonal Antibody (Product # PA5-80208) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0. 1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5,000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit. A specific band was detected for RENT1/hUPF1 at approximately 140 kDa. The expected band size for RENT1/hUPF1 is at 124 kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of UPF1 was achieved by transfecting U-2 OS with UPF1 specific siRNAs (Silencer® select Product # s11927, s11926). Western blot analysis (Fig. a) was performed using modified whole cell extracts from the UPF1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with UPF1 Polyclonal Antibody (Product # PA5-80208, 0.25µg/ml) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to UPF1. An uncharacterized band was observed at ~70 kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-UPF1 Monoclonal Antibody (Product # PA5-80208) and 124 kDa band corresponding to UPF1 was observed along with an uncharacterized band (*) across the samples tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of U-2 OS (Lane1), PC-3 (Lane2), HEK 293 (Lane 3)and tissue extracts of Mouse testis (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0342BOX). 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 (0.25µg/ml) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005)
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry analysis of RENT1/hUPF1 using anti-RENT1/hUPF1 antibody (Product # PA5-80208) . RENT1/hUPF1 was detected in a section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum and then incubated with 2μg/mL rabbit anti-RENT1/hUPF1 antibody (Product # PA5-80208) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of UPF1 on paraffin-embedded rat intestine tissue. Sample was incubated with UPF1 polyclonal antibody (Product# PA5-80208).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of UPF1 on paraffin-embedded mouse intestine tissue. Sample was incubated with UPF1 polyclonal antibody (Product# PA5-80208).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of UPF1 on paraffin-embedded human intestinal cancer tissue. Sample was incubated with UPF1 polyclonal antibody (Product# PA5-80208).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometry of UPF1 in PC-3 cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with UPF1 Polyclonal Antibody (Product # PA5-80208) at a dilution of 1 μg (per 1x10^6 cells), followed by DyLight®488 conjugated goat anti-rabbit IgG (for 30 minutes at 20°C) using 5-10 μg (per 1x10^6 cells) dilution.