Anti-EVI1/MECOM Antibody Picoband™

Product Name

Anti-EVI1/MECOM Antibody Picoband™

View all EVI-1 Antibodies

SKU/Catalog Number

A04590-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-EVI1/MECOM Antibody Picoband™ catalog # A04590-1. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human.

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-EVI1/MECOM Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A04590-1)

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 EVI1/MECOM recombinant protein (Position: E125-D743).

*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

A04590-1 is reactive to MECOM in Human

Applications

A04590-1 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

138 kDa

Calculated molecular weight

Background of EVI-1

MDS1 and EVI1 complex locus protein EVI1 (MECOM) also known as ecotropic virus integration site 1 protein homolog (EVI-1) or positive regulatory domain zinc finger protein 3 (PRDM3) is a protein that in humans is encoded by the MECOM gene. The protein encoded by this gene is a transcriptional regulator and oncoprotein that may be involved in hematopoiesis, apoptosis, development, and cell differentiation and proliferation. The encoded protein can interact with CTBP1, SMAD3, CREBBP, KAT2B, MAPK8, and MAPK9. This gene can undergo translocation with the AML1 gene, resulting in overexpression of this gene and the onset of leukemia. Several transcript variants encoding a few different isoforms have been found 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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Reconstitution

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
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of EVI1/MECOM using anti-EVI1/MECOM antibody (A04590-1).
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 Skov3 whole cell lysates,
Lane 2: human Hel whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human Caco-2 whole cell 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-EVI1/MECOM antigen affinity purified polyclonal antibody (Catalog # A04590-1) 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 EVI1/MECOM at approximately 138 kDa. The expected band size for EVI1/MECOM is at 138 kDa.

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Figure 2. IF analysis of EVI1/MECOM using anti-EVI1/MECOM antibody (A04590-1).
EVI1/MECOM was detected in an immunocytochemical section of A431 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 5 μg/mL rabbit anti-EVI1/MECOM Antibody (A04590-1) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) 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 3. Flow Cytometry analysis of K562 cells using anti-EVI1/MECOM antibody (A04590-1).
Overlay histogram showing K562 cells stained with A04590-1 (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-EVI1/MECOM Antibody (A04590-1, 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

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

Alternative Names

AML1-EVI-1 fusion protein; AML1-EVI-1; ecotropic viral integration site 1; Ecotropic virus integration site 1 protein homolog; EVI1; EVI-1; EVI1MDS1-EVI1; MDS1 and EVI1 complex locus protein EVI1; MDS1 and EVI1 complex locus protein MDS1; MDS1 and EVI1 complex locus; MDS1; MECOM; MGC163392; MGC97004; Myelodysplasia syndrome 1 protein; myelodysplasia syndrome 1; Myelodysplasia syndrome-associated protein 1; oncogene EVI1; PRDM3; zinc finger protein Evi1 MECOM AML1-EVI-1, EVI1, KMT8E, MDS1, MDS1-EVI1, PRDM3, RUSAT2 MDS1 and EVI1 complex locus histone-lysine N-methyltransferase MECOM|AML1-EVI-1 fusion protein|MDS1 and EVI1 complex locus protein EVI1|MDS1 and EVI1 complex locus protein MDS1|PR domain 3|ecotropic virus integration site 1 protein homolog|myelodysplasia syndrome-associated protein 1|oncogene EVI1|zinc finger protein Evi1

For more info on MECOM, check out the MECOM Infographic

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In this infographic, you will see the following information for MECOM: 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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