Anti-LYRIC Antibody Picoband™ (monoclonal, 2E5G3)

Product Name

Anti-LYRIC Antibody Picoband™ (monoclonal, 2E5G3)

View all LYRIC Antibodies

SKU/Catalog Number

M04060-2

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-LYRIC Antibody Picoband™ (monoclonal, 2E5G3) catalog # M04060-2. Tested in Flow Cytometry, IHC, 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-LYRIC Antibody Picoband™ (monoclonal, 2E5G3) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M04060-2)

Host

Mouse

Contents

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

Clonality

Monoclonal

Clone Number

2E5G3

Isotype

Mouse IgG2b

Immunogen

E.coli-derived human LYRIC recombinant protein (Position: D101-Q270). Human LYRIC shares 94% amino acid (aa) sequence identity with both mouse and rat LYRIC.

*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

M04060-2 is reactive to Mtdh in Human

Applications

M04060-2 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

85 kDa

Calculated molecular weight

63.846kDa

Background of LYRIC

MTDH (Metadherin), also known as protein LYRIC or astrocyte elevated gene-1 protein (AEG-1) is a protein that in humans is encoded by the MTDH gene. AEG-1 is involved in HIF-1alpha mediated angiogenesis. AEG-1 also interacts with SND1 and involved in RNA-induced silencing complex (RISC) and plays very important role in RISC and miRNA functions. AEG-1 induces an oncogene called Late SV40 factor (LSF/TFCP2) which is involved in thymidylate synthase (TS) induction and DNA biosynthesis synthesis. Late SV40 factor (LSF/TFCP2) enhances angiogenesis by transcriptionally up-regulating matrix metalloproteinase-9 (MMP9). AEG-1 acts as an oncogene in melanoma, malignant glioma, breast cancer and hepatocellular carcinoma. It is highly expressed in these cancers and helps in progression and development of these cancers. It is induced by c-Myc oncogene and plays very important role in anchorage independent growth of cancer cells.

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
Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human
Flow Cytometry, 1-3 μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of LYRIC using anti-LYRIC antibody (M04060-2).
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 U-87MG whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human PC-3 whole cell lysates,
Lane 4: human HepG2 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 mouse anti-LYRIC antigen affinity purified monoclonal antibody (Catalog # M04060-2) 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for LYRIC at approximately 85 kDa. The expected band size for LYRIC is at 75 kDa.

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Figure 2. IHC analysis of LYRIC using anti-LYRIC antibody (M04060-2).
LYRIC was detected in a paraffin-embedded section of human colorectal adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml mouse anti-LYRIC Antibody (M04060-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.

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Figure 3. Flow Cytometry analysis of U87 cells using anti-LYRIC antibody (M04060-2).
Overlay histogram showing U87 cells stained with M04060-2 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-LYRIC Antibody (M04060-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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

Alternative Names

3D3; AEG1; AEG-13D3/LYRIC; AEG1LYRIC/3D3; Astrocyte elevated gene-1 protein; LYRIC; LYRICMetastasis adhesion protein; Lysine-rich CEACAM1 co-isolated protein; Metadherin; MTDH; protein LYRIC Mtdh|2610103J23Rik, 3D3, 3D3/Ly, 3D3/Lyric, AEG-, AEG-1, AV353288, D8Bwg1112e, Ly, Lyric|metadherin|protein LYRIC|lysine-rich CEACAM1 co-isolated protein|metastasis adhesion protein

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