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 (Paraffin-embedded Sections) (IHC (p)), Immunocytochemistry (ICC)

Purpose: Rabbit IgG polyclonal antibody for Nucleoside diphosphate kinase B(NME2) detection. Tested with WB, IHC-P, ICC in Human,Mouse,Rat.

Sequence: EELVDYKSCA HDWVYE

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

Characteristics: Rabbit IgG polyclonal antibody for Nucleoside diphosphate kinase B(NME2) detection. Tested with WB, IHC-P, ICC in Human,Mouse,Rat.
Gene Name: NME/NM23 nucleoside diphosphate kinase 2
Protein Name: Nucleoside diphosphate kinase B(NDK B)

Purification: Immunogen affinity purified.

Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human NME2(137-152aa EELVDYKSCAHDWVYE), different from the related mouse and rat sequences by one amino acid.

Isotype: IgG

Application Details

Application Notes: WB: Concentration: 0.1-0.5 μg/mL, Tested Species: Human, Rat, Predicted Species: Mouse
IHC-P: Concentration: 0.5-1 μg/mL, Tested Species: Human, Mouse, Rat, Epitope Retrieval by Heat: Boiling the paraffin sections in 10 mM citrate buffer, pH 6.0, for 20 mins is required for the staining of formalin/paraffin sections.
ICC: Concentration: 0.5-1 μg/mL, Tested Species: Human
Notes: Tested Species: Species with positive results. Predicted Species: Species predicted to be fit for the product based on sequence similarities. Other applications have not been tested. Optimal dilutions should be determined by end users.

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 Sodium azide and Thimerosal (Merthiolate): 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: At -20°C for one year. After reconstitution, at 4°C for one month.
It can also be aliquotted and stored frozen at -20 °C for a longer time. Avoid repeated freezing and thawing.

Expiry Date: 12 months

Target Details for NME2

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

Alternative Name: NME2 (NME2 Products)

Synonyms: NDKB antibody, NDPK-B antibody, NDPKB antibody, NM23-H2 antibody, NM23B antibody, PUF antibody, nm23-2 antibody, p18-12d antibody, nm23-M2 antibody, NM23-C2 antibody, NME/NM23 nucleoside diphosphate kinase 2 antibody, non-metastatic cells 2, protein (NM23B) expressed in antibody, NME2 antibody, Nme2 antibody

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.

Synonyms: C myc purine binding transcription factor PUF antibody|C myc transcription factor antibody|C-myc purine-binding transcription factor PUF antibody|Histidine protein kinase NDKB antibody|MGC111212 antibody|MGC2212 antibody|NDK B antibody|NDKB antibody|NDKB_HUMAN antibody|NDP kinase B antibody|NDPK B antibody|NDPKB antibody|NM23 H2 antibody|nm23-H2 antibody|NM23B antibody|nme2 antibody|Non metastatic cells 2, protein(NM23B) expressed in antibody|Nucleoside diphosphate kinase B antibody|Nucleotide diphosphate kinase B antibody|PUF antibody

UniProt: P22392