Antibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
- Immunocytochemistry [2]
- Flow cytometry [2]
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Validation data
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- Product number
- PA5-114371 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- NUP214 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 500 µg/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)
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- Experimental details
- Western blot analysis of NUP214 in the following samples: Lane 1: human HeLA whole cell lysates, Lane 2: human HEK293 whole cell lysates, Lane 3: human K562 whole cell lysates, Lane 4: human THP-1 whole cell lysates. Samples consisting of 50 µg (reducing conditions) of protein was separated with 5-20% SDS-PAGE gel (70V, Stacking gel; 90V, Resolving gel; 2-3 hrs.), transferred to a Nitrocellulose membrane (150mA, 50-90 min) and washed with TBS-0.1% Tween (3 times, 5 minutes/wash) and blocked with 5% Non-fat Milk/TBS (1.5 hrs., room temperature). The membrane was incubated in NUP214 polyclonal antibody (Product # PA5-114371) at a dilution of 0.5 µg/mL (overnight, 4°C), followed by goat anti-rabbit IgG-HRP and chemiluminescence (ECL) with a dilution of 1:10,000 (1.5 hours, room temperature).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of NUP214 in, Lane 1: mouse spleen tissue lysates, Lane 2: mouse thymus tissue 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 50 µg of sample under reducing conditions. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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 NUP214 Polyclonal Antibody (Product # PA5-114371) 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 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit. A specific band was detected for NUP214 at approximately 280KD. The expected band size for NUP214 is at 214KD.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry analysis of NUP214 in A431 cells. Antigen retrieval was performed with enzyme antigen retrieval (15 min). Samples were blocked with 10% goat serum and incubated in NUP214 polyclonal antibody (Product # PA5-114371) at a dilution of 2 µg/mL (overnight, 4°C), followed by DyLight 488 conjugated goat anti-rabbit IgG (30 min, 37°C) and DAPI at a dilution of 1:100.
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunocytochemistry analysis of NUP214 using anti-NUP214 antibody (Product # PA5-114371). NUP214 was detected in a section of NIH/3T3 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-NUP214 antibody (Product # PA5-114371) overnight at 4°C. DyLight®594 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
- Flow Cytometry of NUP214 in HL-60 cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with NUP214 Polyclonal Antibody (Product # PA5-114371) 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometry of NUP214 in HEPA1-6 cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with NUP214 Polyclonal Antibody (Product # PA5-114371) 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.