PA5-28922
antibody from Invitrogen Antibodies
Targeting: DKC1
Cbf5, DKC, dyskerin, NAP57, NOLA4, XAP101
Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA5-28922 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- DKC1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: A549, HeLa, HepG2, HCT116. Predicted reactivity: Mouse (89%), Rat (80%), Zebrafish (82%), Pig (84%), Chicken (80%), Sheep (90%), Rhesus Monkey (98%), Bovine (92%). 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 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 using DKC1 Polyclonal Antibody (Product # PA5-28922). Sample (30 µg of whole cell lysate). Lane A: HeLa. Lane B: HepG2 . 7.5% SDS PAGE. DKC1 Polyclonal Antibody (Product # PA5-28922) diluted at 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of DKC1 was achieved by transfecting Hep G2 with DKC1 specific (Silencer® select Product # s4110, s4111). Western blot analysis (Fig. a) was performed using Modified whole cell extracts (1% SDS) from the untransfected cells (Lane 1), non-specific scrambled siRNA transfected cells (Lane 2) and DKC1 knockdown cells (Lane 3). The blot was probed with DKC1 Polyclonal Antibody (Product # PA5-28922, 1:1000 dilution) 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). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to DKC1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using DKC1 Polyclonal Antibody (Product # PA5-28922) and a 57 kDa band corresponding to DKC1 was observed in cell lines tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of MCF7 (Lane 1) and Hep G2 (Lane 2) 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).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of DKC1 in paraformaldehyde-fixed HeLa cells using a DKC1 polyclonal antibody (Product # PA5-28922) (Green) at a 1:500 dilution. Alpha-tubulin filaments were labeled with Product # PA5-29281 (Red) at a 1:2000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of DKC1 was performed using 70% confluent log phase Hep G2 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 DKC1 Polyclonal Antibody (Product # PA-28922) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing localization to nucleus. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded BT474 xenograft, using DKC1 (Product # PA5-28922) antibody at 1:250 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.