Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [1]
- Other assay [1]
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Validation data
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- Product number
- PA5-76170 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- KAP3 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen and the purity is > 95% (by SDS-PAGE).
- Reactivity
- Human, Mouse, Rat
- 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 analysis of KAP3. Samples were incubated with KAP3 polyclonal antibody (Product # PA5-76170).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of KAP3. Samples were incubated with KAP3 polyclonal antibody (Product # PA5-76170).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- KD of KAP3 was achieved by transfecting SH-SY5Y with KAP3 specific siRNAs (Silencer® select Product # S22677, S22676). Western blot analysis (Fig. a) was performed using whole cell extracts from the KAP3 KD cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with KAP3 Polyclonal Antibody (Product # PA5-76170, 1:500 dilution) and Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (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 KAP3.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-KAP3 Polyclonal Antibody (Product # PA5-76170) and a 80 kDa band corresponding to KAP3 was observed across cell lines and tissue tested. Whole cell extracts (30 µg lysate) of A-431 (Lane 1), NIH/3T3 (Lane 2), A549 (Lane 3), SH-SY5Y (Lane 4), DU 145 (Lane 5) and tissue extract (30 µg lysate) of Mouse Brain (Lane 6) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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 (1:500 dilution) and detected by chemiluminescence 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). An uncharacterized band at ~32 kDa was also observed in some of the samples tested.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of KAP3 was performed using 70% confluent log phase HeLa 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 KAP3 Polyclonal Antibody (Product # PA5-76170) at 5µg/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 staining in cytosol and nucleus. Panel e represents 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
- Immunoprecipitation of KAP3 in extracts of HeLa cells (200 µg). Samples were incubated with 3 µg of KAP3 polyclonal antibody (Product # PA5-76170).