Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [3]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA5-20388 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Bub3 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- A suggested positive control is Jurkat cell lysate. PA5-20388 can be used with blocking peptide PEP-0505.
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 1 mg/mL
- Storage
- Maintain refrigerated at 2-8°C for up to 3 months. For long term storage store at -20°C
Submitted references Identification of host proteins differentially associated with HIV-1 RNA splice variants.
Knoener R, Evans E 3rd, Becker JT, Scalf M, Benner B, Sherer NM, Smith LM
eLife 2021 Feb 25;10
eLife 2021 Feb 25;10
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Jurkat cell lysate using a Bub3 polyclonal antibody (Product # PA5-20388) at (A) 0.5 and (B) 1 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of bub3 in Jurkat cell lysate with Bub3 Polyclonal Antibody (Product # PA5-20388) at (A) 0.5 and (B) 1 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- KD of Bub3 was achieved by transfecting HeLa with Bub3 specific siRNAs (Silencer® select Product # s17556, s2451). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the Bub3 KD cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Bub3 Polyclonal Antibody (Product # PA5-20388, 0.5 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 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 Bub3. Certain uncharacterized bands at ~60 and ~110 kDa were observed across the samples..
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Bub3 Polyclonal Antibody (Product # PA5-20388) and 37 kDa band corresponding to Bub3 was observed across cell lines and tissues tested along with certain uncharacterized bands at ~60 and ~110 kDa. Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), A549 (Lane 2), A-431 (Lane 3), Hep G2 (Lane 4), PC-3 (Lane 5), HEK-293 (Lane 6), Jurkat (Lane 7), SK-BR-3 (Lane 8), tissue extracts of Mouse Brain (Lane 9) and Mouse Spleen (Lane 10) were electrophoresed using Novex® 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 (0.5 µg/mL) 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).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Jurkat cells using a Bub3 polyclonal antibody (Product # PA5-20388) at a 20 µg/mL dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry of Bub3 in Jurkat cells with Bub3 Polyclonal Antibody (Product # PA5-20388) at 10 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence of bub3 in Jurkat cells with Bub3 Polyclonal Antibody (Product # PA5-20388) at 20 µg/mL.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry staining of Jurkat cells using a Bub3 polyclonal antibody (Product # PA5-20388) at a 10 µg/mL dilution.