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Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [5]
- Immunocytochemistry [1]
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- Product number
- PA5-40122 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- DGCR8 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Recommended positive controls: 293T, A431, HeLa, and HepG2 cells. Predicted reactivity: Mouse (82%), Rat (80%), Rhesus Monkey (100%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 1.15 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 DGCR8 in various whole cell extracts (30 µg). Samples were separated by 7.5% SDS-PAGE and the membrane was probed with a DGCR8 polyclonal antibody (Product # PA5-40122) at a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- DGCR8 Polyclonal Antibody detects DGCR8 protein by western blot analysis. Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with DGCR8 Polyclonal Antibody (Product # PA5-40122) diluted at a dilution of 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-DGCR8 Polyclonal Antibody (Product # PA5-40122) and a 110 kDa band corresponding to DGCR8 was observed across the cell lines tested. Whole cell extracts (1%SDS) (30 µg lysate) of A-431 (Lane 1), A549 (Lane 2), HeLa (Lane 3), MCF7 (Lane 4), K-562 (Lane 5) and HEK-293 (Lane 6) were electrophoresed using NuPAGE® 4-12 % Bis-Tris 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 (1:1000 dilution) 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).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- DGCR8 Polyclonal Antibody detects DGCR8 protein by western blot analysis. Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with DGCR8 Polyclonal Antibody (Product # PA5-40122) diluted at a dilution of 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of DGCR8 was achieved by transfecting HEK-293 cells with DGCR8 specific siRNAs (Silencer® select Product # s29062, s29061). Western blot analysis (Fig. a) was performed using whole cell extracts from the DGCR8 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with DGCR8 Antibody (Product # PA5-40122, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 0.25µg/ml, 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 DGCR8.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of DGCR8 was performed using HEK-293 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 DGCR8 Polyclonal Antibody (Product # PA5-40122) at 1:100 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 conjugate (Product # A32790) at a dilution of 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 composite image showing nuclear localization of DGCR8 in HEK-293 cells. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
