Anti-MBD4/MED1 Antibody Picoband™

Product Name

Anti-MBD4/MED1 Antibody Picoband™

View all MBD4 Antibodies

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.

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

Applications

A03462 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

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.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

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, 1-3μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5μg/ml, Human

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).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 (Red line) was also used as a control.

Gene/Protein Information For MBD4 (Source: Uniprot.org, NCBI)

Alternative Names

EC 3.2.2.-; G/T mismatch glycosylase; G/U mismatch glycosylase; MBD4; MED1G/5-fluorouracil mismatch glycosylase with biphasic kinetics; methyl-CpG binding domain protein 4; methyl-CpG-binding domain protein 43,N(4)-ethenocytosine glycosylase; Methyl-CpG-binding endonuclease 1; Methyl-CpG-binding protein MBD4; Mismatch-specific DNA N-glycosylase; putative methyl-CpG binding protein 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

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