Anti-DNMT3B Antibody Picoband™

Product Name

Anti-DNMT3B Antibody Picoband™

View all Dynamin 2 Antibodies

SKU/Catalog Number

A00319-5

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-DNMT3B Antibody Picoband™ catalog # A00319-5. Tested in ELISA, Flow Cytometry, WB applications. This antibody reacts with Human, Mouse, Rat, Monkey.

Storage & Handling

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 freezing and thawing.

Cite This Product

Anti-DNMT3B Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00319-5)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human DNMT3B recombinant protein (Position: E271-E853).

*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

A00319-5 is reactive to DNM2 in Human, Monkey, Mouse, Rat

Applications

A00319-5 is guaranteed for ELISA, Flow Cytometry, WB Boster Guarantee

Observed Molecular Weight

100 kDa

Calculated molecular weight

98.064kDa

Background of Dynamin 2

DNA (cytosine-5-)-methyltransferase 3 beta, is an enzyme that in humans in encoded by the DNMT3B gene. CpG methylation is an epigenetic modification that is important for embryonic development, imprinting, and X-chromosome inactivation. Studies in mice have demonstrated that DNA methylation is required for mammalian development. This gene encodes a DNA methyltransferase which is thought to function in de novo methylation, rather than maintenance methylation. The protein localizes primarily to the nucleus and its expression is developmentally regulated. Mutations in this gene cause the immunodeficiency-centromeric instability-facial anomalies (ICF) syndrome. Eight alternatively spliced transcript variants have been described. The full length sequences of variants 4 and 5 have not been determined.

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

Adding 0.2 ml of distilled water will yield a concentration of 500 μg/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, 0.25-0.5 μg/ml, Human, Monkey, Mouse, Rat
Flow Cytometry, 1-3 μg/1x10⁶ cells, Human, Mouse, Rat
Direct ELISA, 0.1-0.5 μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of DNMT3B using anti-DNMT3B antibody (A00319-5).
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 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: monkey COS-7 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: rat heart tissue lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: mouse heart tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-DNMT3B antigen affinity purified polyclonal antibody (Catalog # A00319-5) 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 DNMT3B at approximately 100 kDa. The expected band size for DNMT3B is at 96 kDa.

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Figure 2. Flow Cytometry analysis of CACO-2 cells using anti-DNMT3B antibody (A00319-5).
Overlay histogram showing CACO-2 cells stained with A00319-5 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DNMT3B Antibody (A00319-5, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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Figure 3. Flow Cytometry analysis of ANA-1 cells using anti-DNMT3B antibody (A00319-5).
Overlay histogram showing ANA-1 cells stained with A00319-5 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DNMT3B Antibody (A00319-5, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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Figure 4. Flow Cytometry analysis of C6 cells using anti-DNMT3B antibody (A00319-5).
Overlay histogram showing C6 cells stained with A00319-5 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DNMT3B Antibody (A00319-5, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For DNM2 (Source: Uniprot.org, NCBI)

Alternative Names

CMTDI1CMT2M; CMTDIBdynamin II; DI-CMTB; DYN2EC 3.6.5.5; dynamin 2; dynamin-2; DYNII DNM2 CMT2M, CMTDI1, CMTDIB, DI-CMTB, DYN2, DYNII, LCCS5 dynamin 2 dynamin-2|dynamin II

For more info on DNM2, check out the DNM2 Infographic

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In this infographic, you will see the following information for DNM2: 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].

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