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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
- Immunocytochemistry [2]
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- Product number
- PA5-65266 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- FANCI Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Immunogen sequence: YYLQVCQSSGG IPKNMEKLVK LSGSHLTPLC YSFISYVQNK SKSLNYTGEK KEKPAAVATA MARVLRETKP IPNLIFAIEQ YEKFLIHLSK KSKVNLMQHM KLSTSRDFKI KGN Highest antigen sequence identity to the following orthologs - mouse 81%, rat 82%.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.2 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
- Knockdown of KIAA1794 was achieved by transfecting A-431 cells with KIAA1794 specific siRNAs (Silencer® select Product # s226830, s226829). Western blot analysis (Fig. a) was performed using whole cell extracts from the KIAA1794 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed KIAA1794 Polyclonal Antibody (Product # PA5-65266, 0.5 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 ug/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 KIAA1794.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-KIAA1794 Polyclonal Antibody (Product # PA5-65266) and a 160kDa band corresponding to KIAA1794 was observed in Jurkat, MOLT-4, MCF7, A-431, HEK-293, HeLa, K-562, Daudi and U-87 MG. Whole cell extracts (30 µg lysate) of Jurkat (Lane 1), MOLT-4 (Lane 2), MCF7 (Lane 3), A-431 (Lane 4), HEK-293 (Lane 5), HeLa (Lane 6), K-562 (Lane 7), Daudi (Lane 8) and U-87 MG (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 (0.5 µg/mL) and detected by chemiluminescence with Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (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 staining of FANCI in human cell line U-251 MG shows localization to nucleoplasm and cytosol. Samples were probed using a FANCI Polyclonal Antibody (Product # PA5-65266).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of KIAA1794 was performed using Jurkat 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 KIAA1794 Polyclonal Antibody (Product # PA5-65266) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) 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 merged image showing predominant nuclear as well as cytosolic localization. Panel e represents cells with no primary antibody to assess background. The images were captured at 60X magnification.
