Explore
Validate
Learn
Western blotAntibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- ELISA [2]
- Chromatin Immunoprecipitation [2]
Submit
Validation data
Reference
Comment
Report error
- Product number
- 49-1027 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- MBD1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Other
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 50 µg
- Concentration
- 0.7 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Nuclear extracts of HeLa cells (40 μg) were analysed by Western blot using the anti-MBD1 antibody (Product # 49-1027) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right, the marker (in kDa) is shown on the left.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Titer determination. To determine the titer, an ELISA was performed using anti-MBD1 crude serum, purified anti-MBD1 antibodies (Product # 49-1027), and the column flow through obtained from the antibody purification step. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1:16, 500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- To determine the titer of the antibody, an ELISA was performed using a serial dilution of the anti-human MBD1 antibody (Product # 49-1027), crude serum and Flow Through. The plates were coated with the peptide used for immunization of the rabbit. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1:20,000.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- ChIP assays were performed using human osteosarcoma (U-2 OS) cells, the anti-MBD1 antibody (Product # 49-1027), and optimized PCR primer sets. Each ChIP assay used sheared chromatin from 1 million cells and 1.7 µg of MBD1 antibody or beads only. This figure shows the recovery as a percentage of the input DNA. Left: Recovery by MBD1 or beads only of the MLH1 promoter, which specifically binds MBD1. Right: Recovery of the CDC6 promoter (used as a negative control) by MBD1 or beads only.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- ChIP assays were performed using human osteosarcoma (U2OS) cells, the anti-MBD1 antibody (Product # 49-1027) and optimized PCR primer sets. Sheared chromatin from 1x10e6 cells and 1.5 μg of antibody were used per ChIP experiment. Beads only were used as a negative IP control. Quantitative PCR was performed with primers for the promoters of the MLH1 gene (used as a positive control) and CDC6 gene (used as a negative control). The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
