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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Chromatin Immunoprecipitation [1]
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- Product number
- 703489 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- BRPF1 Recombinant Rabbit Monoclonal Antibody (19H16L8)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- This antibody is predicted to react with Mouse, Rat, Bovine.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- 19H16L8
- Vial size
- 100 µg
- Concentration
- 0.5 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 was performed using Anti-BRPF1 Recombinant Rabbit Monoclonal Antibody (Product # 703489) and 160, 150, 140 kDa bands corresponding to isoforms of BRPF1 (PMID: 18794358) was observed across the cell lines tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of U-2 OS (Lane 1), HeLa (Lane 2), HEK-293 (Lane 3), HT-1080 (Lane 4), NTERA-2 (Lane 5) and PC-3 (Lane 6) were electrophoresed using Novex® NuPAGE® 3-8 % Tris-Acetate gel (Product # EA0375BOX). Resolved proteins were then transferred onto a nitrocellulose membrane by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution) using the iBright FL 1500 (Product # A44115). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- For immunofluorescence analysis, HeLa cells were fixed and permeabilized for detection of endogenous BRPF1 using Anti-BRPF1 Recombinant Rabbit Monoclonal Antibody (Product # 703489, 1:100 dilution) and labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 conjugate (Product # A32731, 1:2000). Panel a) shows representative cells that were stained for detection and localization of BRPF1 protein (green), Panel b) is stained for nuclei (blue) using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415, 1:1000). Panel d) is a composite image of Panels a, b and c clearly demonstrating strong nuclear and faint cytoplasmic localization of BRPF1. Panel e) represents control 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
- Chromatin Immunoprecipitation (ChIP) assay of endogenous BRPF1 protein using BRPF1 Antibody: ChIP was performed using Anti-BRPF1 Recombinant Rabbit Monoclonal Antibody (SKU # 703489, 5 µg) on sheared chromatin from 2 million HeLa cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to promoter regions of RUNX2, PIK3CA genes and SAT2 satellite repeats. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.
