Anti-CD10/MME Antibody Picoband™

Product Name

Anti-CD10/MME Antibody Picoband™

View all Neprilysin/CD10 Antibodies

SKU/Catalog Number

PB9058

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-CD10/MME Antibody Picoband™ catalog # PB9058. Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Human, 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-CD10/MME Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9058)

Host

Rabbit

Contents

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

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human CD10 recombinant protein (Position: Y52-W750). Human CD10 shares 94% amino acid (aa) sequences identity with both mouse and rat CD10.

*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

PB9058 is reactive to MME in Human, Rat

Applications

PB9058 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

100 kDa

Calculated molecular weight

85.514kDa

Background of Neprilysin/CD10

CD10, also known as membrane metallo-endopeptidase, neutral endopeptidase (NEP), Neprilysin, or common acute lymphoblastic leukemia antigen (CALLA), is a zinc-dependent metalloprotease enzyme that degrades a number of small secreted peptides, most notably theamyloid beta peptide whose abnormal misfolding and aggregation in neural tissue has been implicated as a cause of Alzheimer's disease. This gene is localized to human chromosome 3 by study of somatic cell hybrids and regionalized the location to 3q21-q27 by in situ hybridization. By cDNA transfection analysis, CD10 is confirmed as a functional neutral endopeptidase of the type that has previously been called enkephalinase. CD10 has also been called atriopeptidase. Atriopeptidase specifically degrades atrial natriuretic factor. A specific enzyme inhibitor was developed and reported that it had effects similar to those of low-dose ANF infusion. These effects include diuresis, natriuresis, vasodilatation, and suppression of the renin-angiotensin-aldosterone system.

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, 0.1-0.5μg/ml, Human, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Flow Cytometry, 1-3 μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of CD10/MME using anti-CD10/MME antibody (PB9058).
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 Daudi whole cell lysates,
Lane 2: human U-87MG whole cell lysates,
Lane 3: human placenta tissue lysates,
Lane 4: rat kidney 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-CD10/MME antigen affinity purified polyclonal antibody (Catalog # PB9058) 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 CD10/MME at approximately 100 kDa. The expected band size for CD10/MME is at 85 kDa.

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Figure 2. IHC analysis of CD10/MME using anti-CD10/MME antibody (PB9058).
CD10/MME was detected in a paraffin-embedded section of human lymphoma 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 rabbit anti-CD10/MME Antibody (PB9058) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 3. IHC analysis of CD10/MME using anti-CD10/MME antibody (PB9058).
CD10/MME was detected in a paraffin-embedded section of human placenta 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 rabbit anti-CD10/MME Antibody (PB9058) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 4. IHC analysis of CD10/MME using anti-CD10/MME antibody (PB9058).
CD10/MME was detected in a paraffin-embedded section of human renal cell carcinoma 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 rabbit anti-CD10/MME Antibody (PB9058) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 5. Flow Cytometry analysis of Daudi cells using anti-CD10/MME antibody (PB9058).
Overlay histogram showing Daudi cells stained with PB9058 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CD10/MME Antibody (PB9058, 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 MME (Source: Uniprot.org, NCBI)

Alternative Names

Atriopeptidase; CALLA; CALLAmembrane metallo-endopeptidase (neutral endopeptidase, enkephalinase); CD10 antigen; CD10; CD10); CD10membrane metallo-endopeptidase variant 1; Common acute lymphocytic leukemia antigen; DKFZp686O16152; EC 3.4.24; EC 3.4.24.11; Enkephalinase; EPN; Leu-19; membrane metallo-endopeptidase variant 2; membrane metallo-endopeptidase; MGC126681; MGC126707; MME; NEPmembrane metallo-endopeptidase (neutral endopeptidase, enkephalinase, CALLA; Neprilysin; Neutral Endopeptidase 24.11; Neutral endopeptidase; NKH1; SFE; Skin fibroblast elastase MME CALLA, CD10, CMT2T, NEP, SCA43, SFE membrane metalloendopeptidase neprilysin|atriopeptidase|common acute lymphocytic leukemia antigen|membrane metallo-endopeptidase (neutral endopeptidase, enkephalinase, CALLA, CD10)|membrane metallo-endopeptidase variant 1|membrane metallo-endopeptidase variant 2|neprilysin-390|neprilysin-411|neutral endopeptidase 24.11|skin fibroblast elastase

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