NME2 antibody (C-Term)

Catalog No. ABIN3044198

Product Details anti-NME2 Antibody

Target: NME2 (Nucleoside diphosphate kinase B (NME2))

Binding Specificity: AA 137-152, C-Term

Reactivity: Human, Mouse, Rat

Host: Rabbit

Clonality: Polyclonal

Conjugate: This NME2 antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), Immunocytochemistry (ICC), Flow Cytometry (FACS)

Purpose: Anti-NME2 Antibody Picoband®

Sequence: EELVDYKSCA HDWVYE

Cross-Reactivity (Details): No cross-reactivity with other proteins

Characteristics: Anti-NME2 Antibody (ABIN3044198). Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.

Purification: Immunogen affinity purified.

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.

Isotype: IgG

Application Details

Application Notes: Western blot, 0.1-0.5 μg/mL, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1 μg/mL, Human, Mouse, Rat
Immunocytochemistry , 0.5-1 μg/mL, Human
Immunocytochemistry/Immunofluorescence, 2 μg/mL, Human
Flow Cytometry (Fixed), 1-3 μg/1x10⁶ cells, Human1. Backer, J. M., Mendola, C. E., Kovesdi, I., Fairhurst, J. L., O'Hara, B., Eddy, R. L., Jr., Shows, T. B., Mathew, S., Murty, V. V. V. S., Chaganti, R. S. K. Chromosomal localization and nucleoside diphosphate kinase activity of human metastasis-suppressor genes NM23-1 and NM23-2.Oncogene 8: 497-502, 1993. 2. Masse, K., Dabernat, S., Bourbon, P.-M., Larou, M., Amrein, L., Barraud, P., Perel, Y., Camara, M., Landry, M., Lacombe, M.-L., Daniel, J.-Y.Characterization of the nm23-M2, nm23-M3 and nm23-M4 mouse genes: comparison with their human homologs. Gene 296: 87-97, 2002. 3. Srivastava, S., Li, Z., Ko, K., Choudhury, P., Albaqumi, M., Johnson, A. K., Yan, Y., Backer, J. M., Unutmaz, D., Coetzee, W. A., Skolnik, E. Y.Histidine phosphorylation of the potassium channel KCa3.1 by nucleoside diphosphate kinase B is required for activation of KCa3.1 and CD4 T cells. Molec. Cell 24: 665-675, 2006.

Comment:

Antibody can be supported by chemiluminescence kit ABIN921124 in WB, supported by ABIN921231 in IHC(P) and ICC.

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #300030 (Immunohistochemistry)

by: Human Protein Atlas

No.: #300030

Date: 12/13/2016

Antigen: P18

Method validated: Immunohistochemistry

Positive Control: tonsil, urinary bladder

Notes: Passed. ABIN3044198 staining is consistent with staining pattern of other commercial P18/NME2 antibodies.

Validation Images

Staining of human tonsil (A.) and urinary bladder (B.) sections in a tissue microarray with ABIN3044198. See the protocol for detailed information.

Full Methods

Primary Antibody: ABIN3044198

Secondary Antibody: HRP polymer (ThermoFisher Scientific, TL-125-PH)

Full Protocol:

• Sample preparation:
• • Fixation and TMA preparation according to PMID: 22688270.
  • Cut 4µm TMA sections using a waterfall microtome (ThermoFisher Scientific, HM355S).
  • Dry sections ON at RT and then back them for 12-24h at 50°C.
• Deparaffinization and rehydration in xylene and graded alcohol series:
• • xylene twice for 3min.
  • 100% ethanol for 1min.
  • 95% ethanol for 1min.
  • 0.3% H₂O₂ in 95% ethanol for 5min to block endogenous peroxidase.
  • 70% ethanol for 1min.
  • 50% ethanol for 1min.
  • distilled H₂O for 2min.
• Antigen Retrieval by Heat Induced Epitope Retrieval (HIER):
• • Boil the sections in Lab Vision Citrate buffer pH6.0 (ThermoFisher Scientific, TA-250-PM1X) for 4min at 125°C in a decloaking chamber (Biocare Medical).
  • Allow slides to cool down to 90°C in the decloaking chamber.
• Immunostaining in a Lab Vision Autostainer 480 (ThermorFisher Scientific) at RT with 300µl of reagents for each step:
• • Rinse sections in Lab Vision wash buffer with extra tween added to a final concentration of 0.2% (ThermoFisher Scientific, TA-999-TT and TA-125-TW) wash buffer.
  • Block sections with Lab Vision Ultra V-Block (ThermoFisher Scientific, TA-125-UB) for 5min.
  • Rinse sections 2x in wash buffer.
  • Incubate sections with primary rabbit anti-P18/NME2 antibody (antibodies-online, ABIN3044198) diluted 1:25000 for 30min.
  • Rinse sections 3x in wash buffer.
  • Incubate sections with labeled HRP polymer (ThermoFisher Scientific, TL-125-PH) for 30min.
  • Rinse sections 2x in wash buffer.
  • Develop in DAB Quanto solution (ThermoFisher Scientific, TL-125-QHDX) for 5min.
  • Rinse sections in distilled H₂O.
• Washing, counterstaining, and coverslipping in Autostainer XL (Leica Biosystems, ST5010) at RT:
• • Counterstaining in Mayer’s hematoxylin plus (HistoLab, 01820) for 7.5min.
  • Rinse sections in tap water for 5min.
  • Rinse sections in lithium carbonate water diluted 1:5 for 5min.
  • Dehydration in graded ethanol series and Neo-Clear (Merck Millipore, 109843) according to the manufacturer recommendations.
  • Automated mounting (Leica Biosystems, CV5030) of coverslip with Pertex (Histolab, 00871).
• Microscopy:
• • Image acquisition on a Leica Aperio AT2 and Aperio Scanscope AT at 20x magnification.

Experimental Notes: ABIN3044198 shows staining in most tissues similar to alternative P18/NME2 antibodies. Determination of the appropriate dilution factor proved difficult.

Handling

Format: Lyophilized

Reconstitution: Add 0.2 mL of distilled water will yield a concentration of 500 μg/mL.

Concentration: 500 μg/mL

Buffer: Each vial contains 5 mg BSA, 0.9 mg NaCl, 0.2 mg Na2HPO4, 0.05 mg Thimerosal, 0.05 mg Sodium azide.

Preservative: Thimerosal (Merthiolate), Sodium azide

Precaution of Use: This product contains Thimerosal (Merthiolate) and Sodium azide: POISONOUS AND HAZARDOUS SUBSTANCES which should be handled by trained staff only.

Handling Advice: Avoid repeated freezing and thawing.

Storage: 4 °C,-20 °C

Storage Comment: 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.

Expiry Date: 12 months

Target Details for NME2

Target: NME2 (Nucleoside diphosphate kinase B (NME2))

Alternative Name: NME2 (NME2 Products)

Background:

Synonyms: Nucleoside diphosphate kinase B;NDK B;NDP kinase B;2.7.4.6;C-myc purine-binding transcription factor PUF;Histidine protein kinase NDKB;2.7.13.3;nm23-H2;NME2;NM23B;

Tissue Specificity: Ubiquitously expressed. .

Background: 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.

Sequence Similarities: Belongs to the NDK family.

Molecular Weight: 17 kDa

UniProt: P22392