Anti-NME2 Antibody

Product Name

Anti-NME2 Antibody

View all NM23-H2/NME2 Antibodies

SKU/Catalog Number

PA1830

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-NME2 Antibody catalog # PA1830. Tested in 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-NME2 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1830)

Host

Rabbit

Contents

Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence at the C-terminus of human NME2, different from the related mouse and rat sequences by one amino acid.

*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

PA1830 is reactive to NME2 in Human, Mouse, Rat

Applications

PA1830 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

17 kDa

Calculated molecular weight

17.298kDa

Background of NM23-H2/NME2

NME2 (NME/NM23 nucleoside diphosphate kinase 2) also called non-metastatic cells 2, protein (NM23B) expressed in, NM23-H2, NM23B or NDPKB, is identical to the beta subunit of human erythrocyte NDP kinase. Little to no expression was detected in other mouse tissues examined. Using Northern blot analysis, Masse et al. (2002) detected high expression of mouse Nme2, which they called nm23-M2, in heart, liver, and kidney, with intermediate expression in skeletal muscle. In situ hybridization of 15-day postcoitum mouse embryos showed ubiquitous Nme2 expression. The mouse and human NME2 genes contain 5 exons and span about 6.0 kb. The NME2 gene is mapped on 17q21.33. Srivastava et al. (2006) concluded that histidine phosphorylation regulates KCa3.1 channel activity and that NDPBK is critical to the channel activity and the activation of CD4 T 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

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, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, By Heat
Immunocytochemistry , 0.5-1μg/ml, Human
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Anti-NME2 antibody, PA1830, IHC(P)
IHC(P): Human Intestinal Cancer Tissue

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Anti-NME2 antibody, PA1830, IHC(P)
IHC(P): Rat Cerebellum Tissue

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Anti-NME2 antibody, PA1830, ICC
ICC: HELA Cell

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Figure 1. Western blot analysis of NME2 using anti-NME2 antibody (PA1830).
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: rat heart tissue lysates,
Lane 2: rat brain tissue lysates,
Lane 3: mouse heart tissue lysates,
Lane 4: mouse brain tissue 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-NME2 antigen affinity purified polyclonal antibody (Catalog # PA1830) 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 NME2 at approximately 17KD. The expected band size for NME2 is at 17KD.

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Figure 5. IHC analysis of NME2 using anti-NME2 antibody (PA1830).
NME2 was detected in paraffin-embedded section of mouse intestine tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-NME2 Antibody (PA1830) 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 6. IF analysis of NME2 using anti-NME2 antibody (PA1830).
NME2 was detected in immunocytochemical section of U20S 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-NME2 Antibody (PA1830) overnight at 4°C. DyLight®550 Conjugated Goat Anti-Rabbit IgG (BA1135) 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 7. Flow Cytometry analysis of HL-60 cells using anti-NME2 antibody (PA1830).
Overlay histogram showing HL-60 cells stained with PA1830 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NME2 Antibody (PA1830, 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 NME2 (Source: Uniprot.org, NCBI)

Alternative Names

EC 2.7.13.3; Histidine protein kinase NDKB; MGC2212; NDK B; NDKB; NDP Kinase B2; NDPKB; NDPK-B; NM23B; NM23-H2; NME2; non-metastatic cells 2, protein (NM23B) expressed in; nucleoside diphosphate kinase B; protein (NM23) expressed in NME2 NDKB, NDPK-B, NDPKB, NM23-H2, NM23B, PUF NME/NM23 nucleoside diphosphate kinase 2 nucleoside diphosphate kinase B|HEL-S-155an|NDP kinase B|c-myc purine-binding transcription factor PUF|c-myc transcription factor|epididymis secretory sperm binding protein Li 155an|histidine protein kinase NDKB|non-metastatic cells 2, protein (NM23) expressed in|non-metastatic cells 2, protein (NM23B) expressed in|nucleotide diphosphate kinase B

For more info on NME2, check out the NME2 Infographic

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In this infographic, you will see the following information for NME2: 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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