Antibody data
- Antibody Data
- Antigen structure
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- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [1]
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Validation data
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- Product number
- 720269 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- RAD17 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Other
- Description
- These Polyclonal antibodies are of rabbit origin developed by immunizing animals with proteins or peptides. The polyclonal antibody is purified by affinity purification from the rabbit sera generated after immunizing the rabbits with a specific type of protein or peptide. The purified antibody is tested for its functionality in various relevant research applications. The antibody is developed for Research Use Only and is non-hazardous or non-infectious in nature.
- Concentration
- 0.5 mg/mL
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Supportive validation
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on Nuclear enriched cell extracts (30 µg lysate) of SW480 (Lane 1), K-562 (Lane 2), Raji (Lane 3), HeLa (Lane 4), MCF-7 (Lane 5), MDA-MB-231 (Lane 6) and T-47D (Lane 7) and tissue extracts of Mouse Testis (Lane 8) and Rat Testis (Lane 9). The blots were probed with Anti-Rad17 Rabbit Polyclonal Antibody (Product # 720269, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 77 kDa band corresponding to Rad17 was observed across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- For immunofluorescence analysis, HeLa cells were fixed and permeabilized for detection of endogenous Rad17 using Anti- Rad17 Rabbit Polyclonal Antibody (Product # 720269, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of Rad17 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of Rad17. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.