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Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [4]
- Immunocytochemistry [1]
- Flow cytometry [2]
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- Product number
- MA1-16577 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- BUBR1 Monoclonal Antibody (8G1)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Suggested positive control: Hela whole cell extract, antigen standard for BUB1B (transient overexpression lysate).
- Reactivity
- Human, Mouse, Rat
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 8G1
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- BUBR1 detected in HeLa cell lysate using Product # MA1-16577.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Bub1b in HeLa whole cell extracts using a monoclonal antibody (Product # MA1-16577).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of BUBR1 in HeLa cell lysate. Sample was incubated in BUBR1 monoclonal antibody (Product # MA1-16577).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-BUBR1 Monoclonal Antibody (8G1)(Product # MA1-16577) and a 120kDa band corresponding to BUBR1 was observed across cell lines and tissues tested. Whole cell extracts (30 µg lysate) of RD (Lane 1), SK-O-V3 (Lane 2), HeLa (Lane 3), MDA-MB-231 (Lane 4), MCF7 (Lane 5), PC-3 (Lane 6), Mouse Thymus (Lane 7), Rat Thymus (Lane 8),Mouse Liver (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 (1:1000 Dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).Expression of BUBR1 was found to be higher in all cell lines and tissues except MCF7.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence of an asynchronous cycling population of human cells (U2OS) with MA1-16577. No signal is detected from interphase cells, whereas cells undergoing mitosis accumulate BubR1 at the kinetochores. Image reveals kinetochores at prometaphase.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry of BUBR1 in HeLa cells. Samples were incubated in BUBR1 monoclonal antibody (Product # MA1-16577) using a dilution of 5 µg/mL for 30 minutes at room temperature. Antibody (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Both antibodies were conjugated to Dylight 650.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry of BUBR1 in 1 x 10^6 HEK-293 cells. Samples were incubated in BUBR1 monoclonal antibody (Product # MA1-16577) using a dilution of 1 µg/1x10^6 cells. Antibody (dark blue). Isotype control shown in orange.
