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Product Info Summary
| SKU: | A03462 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-MBD4/MED1 Antibody Picoband®
SKU/Catalog Number
A03462
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-MBD4/MED1 Antibody Picoband® catalog # A03462. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-MBD4/MED1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A03462)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human MBD4/MED1 recombinant protein (Position: A16-S580).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A03462 is reactive to MBD4 in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
66 kDa
Calculated molecular weight
66.051kDa
Background of MBD4
Methyl-CpG-binding domain protein 4 is a protein that in humans is encoded by the MBD4 gene. It is mapped to 3q21.3. The protein encoded by this gene is a member of a family of nuclear proteins related by the presence of a methyl-CpG binding domain (MBD). These proteins are capable of binding specifically to methylated DNA, and some members can also repress transcription from methylated gene promoters. This protein contains an MBD domain at the N-terminus that functions both in binding to methylated DNA and in protein interactions and a C-terminal mismatch-specific glycosylase domain that is involved in DNA repair. Alternatively spliced transcript variants encoding multiple isoforms have been observed for this gene.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A03462 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human A549 whole cell, human Jurkat whole cell, human SK-OV-3 whole cell, human Raji whole cell, human hela whole cell, human A431 whole cell, human MDA-MB-453 whole cell
IHC: mouse brain tissue, rat brain tissue
ICC/IF: Hela cell
FCM: SiHa cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of MBD4/MED1 using anti-MBD4/MED1 antibody (A03462).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human A549 whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: human SK-OV-3 whole cell lysates,
Lane 4: human Raji whole cell lysates,
Lane 5: human hela whole cell lysates,
Lane 6: human A431 whole cell lysates,
Lane 7: human MDA-MB-453 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-MBD4/MED1 antigen affinity purified polyclonal antibody (Catalog # A03462) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for MBD4/MED1 at approximately 66KD. The expected band size for MBD4/MED1 is at 66KD.
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Figure 2. IHC analysis of MBD4/MED1 using anti-MBD4/MED1 antibody (A03462).
MBD4/MED1 was detected in paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-MBD4/MED1 Antibody (A03462) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
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Figure 3. IHC analysis of MBD4/MED1 using anti-MBD4/MED1 antibody (A03462).
MBD4/MED1 was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-MBD4/MED1 Antibody (A03462) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
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Figure 4. IF analysis of MBD4/MED1 using anti-MBD4/MED1 antibody (A03462).
MBD4/MED1 was detected in immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-MBD4/MED1 Antibody (A03462) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Figure 5. Flow Cytometry analysis of SiHa cells using anti-MBD4/MED1 antibody (A03462).
Overlay histogram showing SiHa cells stained with A03462 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-MBD4/MED1 Antibody (A03462, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For MBD4 (Source: Uniprot.org, NCBI)
Gene Name
MBD4
Full Name
Methyl-CpG-binding domain protein 4
Weight
66.051kDa
Alternative Names
Methyl-CpG-binding domain protein 4; Methyl-CpG-binding endonuclease 1; Methyl-CpG-binding protein MBD4; Mismatch-specific DNA N-glycosylase; MBD4; MED1 MBD4 MED1 methyl-CpG binding domain 4, DNA glycosylase methyl-CpG-binding domain protein 4|3,N(4)-ethenocytosine glycosylase|G/5-fluorouracil mismatch glycosylase with biphasic kinetics|G/T mismatch glycosylase|G/U mismatch glycosylase|methyl-CpG binding domain protein 4|methyl-CpG-binding endonuclease 1|methyl-CpG-binding protein MBD4|mismatch-specific DNA N-glycosylase|putative methyl-CpG binding protein
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on MBD4, check out the MBD4 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for MBD4: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-MBD4/MED1 Antibody Picoband® (A03462)
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