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ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
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- Validations
- ELISA [1]
- Chromatin Immunoprecipitation [1]
- Other assay [1]
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- Product number
- 49-1028 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-DNMT3B Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Other
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 5 µg
- Concentration
- Lot Dependent
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Titer determination. To determine, an ELISA was performed using a serial dilution of the anti-DNMT3B antibody (Product # 49-1028) and crude serum in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1:900.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- ChIP assays were performed using human osteosarcoma (U-2 OS) cells, the antibody directed against DNMT3B (Product # 49-1028), and optimized PCR primer sets. Each ChIP assay used sheared chromatin from 1 million cells and 0.2 µg of DNMT3B antibody or beads only. The immunoprecipitated DNA was analysed at two loci: the MLH1 promoter, which is a positive locus for DNMT3B, and the CDC6 promoter, a negative locus for DNMT3B. This figure shows the recovery expressed as a percent of the input DNA. Left: Recovery of the MLH1 promoter by the DNMT3B antibody or beads only. Right: Recovery of the CDC6 promoter by the DNMT3B antibody or beads only.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunoprecipitation of DNMT3B was performed on K562 cells. Antigen-antibody complexes were formed by incubating approximately 500 µg whole cell lysate with 5 µg of DNMT3B polyclonal antibody (Product # 49-1028) rotating 60 min at RT. The immune complexes were captured on 625 µg of anti- rabbit coated Dynabeads (Product # 11204D), washed extensively, and eluted with NuPAGE™ LDS Sample Buffer (Product # NP0007). Samples were resolved onto NuPAGE™ 4-12% Bis-Tris gel (Product # NP0335BOX). Lanes 1 and 3 are input and lanes 2 and 4 are IP. Proteins were transferred to PVDF membrane (Product # IB23001). Membrane was blocked in 5% milk. Target was detected using a DNMT3B polyclonal antibody (Product # 49-1028) at a dilution of 1:2000, followed by a 1:4000 dilution of secondary antibody. Chemiluminescent detection was performed using ECL Western Blotting Substrate (Product # 32106). Data courtesy of the Yeo lab as part of the ENCODE project (www.encodeproject.org).
