Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Flow cytometry [1]
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Validation data
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- Product number
- PA5-95499 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- DYRK1A Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- 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
- 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 DYRK1A in Lane 1: human HeLa whole cell lysates, Lane 2: human MCF-7 whole cell lysates, Lane 3: rat skeletal muscle tissue lysates, Lane 4: mouse skeletal muscle tissue lysates. Electrophoresis was performed with 5-20% SDS-PAGE gel (70V, Stacking gel; 90V Resolving gel, Time: 2-3 hours), transferred to a nitrocellulose membrane and blocked using 5% Non-fat Milk/TBS (1.5 hrs at room temperature). Samples were incubated with DYRK1A polyclonal antibody (Product # PA5-95499) using a 0.5 µg/mL dilution, followed by a goat anti-rabbit IgG-HRP at a dilution of 1:10,000, and developed with enhanced chemiluminescence (ECL).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Dual specificity tyrosine-phosphorylation-regulated kinase 1A was achieved by transfecting SH-SY5Y with Dual specificity tyrosine-phosphorylation-regulated kinase 1A specific siRNAs (Silencer® select Product # S4400, S4399). Western blot analysis (Fig. A) was performed using Nuclear enriched extracts from the Dual specificity tyrosine-phosphorylation-regulated kinase 1A knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with DYRK1A Polyclonal Antibody (Product # PA5-95499, 0.3 µg/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 Dual specificity tyrosine-phosphorylation-regulated kinase 1A.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-DYRK1A Polyclonal Antibody (Product # PA5-95499) and a 90kDa band corresponding to Dual specificity tyrosine-phosphorylation-regulated kinase 1A was observed across all the tested cell lines and tissues, except HEK-293 and Mouse Brain. Nuclear enriched extracts (40 µg lysate) of HEK-293 (Lane 1), K-562 (Lane 2), HEL 92.1.7 (Lane 3), SH-SY5Y (Lane 4), PC-3 (Lane 5), HeLa (Lane 6), Jurkat (Lane 7), Mouse Ovary (Lane 8), Mouse Brain (Lane 9) 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 (0.3 µg/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 SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of DYRK1A in U20S cell. Antigen retrieval was performed on the tissue using IHC enzyme antigen retrieval reagent and blocked with 10% goat serum. Samples were incubated with DYRK1A polyclonal antibody (Product # PA5-95499) at a 2 µg/mL dilution, followed by Cy3 conjugated Goat anti-rabbit IgG using a 1:100 dilution and DAPI.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry of DYRK1A in SiHa cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with DYRK1A polyclonal antibody (Product # PA5-95499) at a dilution of 1 µg (per 1x10^6 cells), followed by 488 conjugated goat anti-rabbit IgG (30 min at 20°C) using a 5-10 µg (per 1x10^6 cells) dilution.