|Validated Species:||Human, Mouse, Rat|
|Application:||Flow Cytometry, IHC, ICC, WB|
Data & Images
|Product Name||Anti-NM23A Antibody|
|Description||Rabbit IgG polyclonal antibody for Nucleoside diphosphate kinase A(NME1) detection. Tested with WB, IHC-P, IHC-F, ICC, FCM in Human;Mouse;Rat.|
|Cite This Product||Anti-NM23A Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1829)|
|Replacement Item||This antibody may replace the following items: sc-136141|sc-14787|sc-14789|sc-14790|sc-166937|sc-17586|sc-17587|sc-28829|sc-343|sc-465|sc-514515|sc-514515-X|sc-56928 from Santa Cruz Biotechnology.|
|Validated Species||Human, Mouse, Rat|
*This antibody is predicted to react with the above species based on antigen sequence similarities. Our Boster Guarantee covers the use of this product with the above species.
|Application||Flow Cytometry, IHC, ICC, WB
*Our Boster Guarantee covers the use of this product in the above tested applications.
**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.
|Recommended Detection Systems||Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P), IHC(F) and ICC.
*Blocking peptide can be purchased at $50. Contact us for more information
**Boster also offers various secondary antibodies for Immunoflourescecne and IHC. Take advantage of the buy 1 primary antibody get 1 secondary antibody for free promotion for the entire year 2018!
|Immunogen||A synthetic peptide corresponding to a sequence at the C-terminus of human NM23A(137-152aa EELVDYTSCAQNWIYE), different from the related mouse sequence by two amino acids and from rat sequence by one amino acid.|
|Cross Reactivity||No cross reactivity with other proteins|
|Contents||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.|
|Concentration||Add 0.2ml of distilled water will yield a concentration of 500ug/ml.|
|Storage||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.|
|Purification||Immunogen affinity purified.|
Protein Target Info (Source: Uniprot.org)
You can check the tissue specificity below for information on selecting positive and negative control.
|Protein Name||Nucleoside diphosphate kinase A(NDK A)|
|Molecular Weight||17149 MW|
|Protein Function||Major role in the synthesis of nucleoside triphosphates other than ATP. The ATP gamma phosphate is transferred to the NDP beta phosphate via a ping-pong mechanism, using a phosphorylated active-site intermediate. Possesses nucleoside-diphosphate kinase, serine/threonine-specific protein kinase, geranyl and farnesyl pyrophosphate kinase, histidine protein kinase and 3'-5' exonuclease activities. Involved in cell proliferation, differentiation and development, signal transduction, G protein- coupled receptor endocytosis, and gene expression. Required for neural development including neural patterning and cell fate determination. During GZMA-mediated cell death, works in concert with TREX1. NME1 nicks one strand of DNA and TREX1 removes bases from the free 3' end to enhance DNA damage and prevent DNA end reannealing and rapid repair. .|
|Tissue Specificity||Isoform 1 is expressed in heart, brain, placenta, lung, liver, skeletal muscle, pancreas, spleen and thymus. Expressed in lung carcinoma cell lines but not in normal lung tissues. Isoform 2 is ubiquitously expressed and its expression is also related to tumor differentiation. Isoform 3 is ubiquitously expressed. .|
|Sequence Similarities||Belongs to the NDK family.|
|Subcellular Localization||Cytoplasm . Nucleus . Cell-cycle dependent nuclear localization which can be induced by interaction with Epstein-barr viral proteins or by degradation of the SET complex by GzmA.|
|Alternative Names||Nucleoside diphosphate kinase A;NDK A;NDP kinase A;126.96.36.199;Granzyme A-activated DNase;GAAD;Metastasis inhibition factor nm23;NM23-H1;Tumor metastatic process-associated protein;NME1;NDPKA, NM23;|
|Research Areas|||epigenetics and nuclear signaling|transcription|cancer susceptibility|tumor suppressors||
Background for Nucleoside diphosphate kinase A(NDK A)
Dilution Ratios/Recommended Concentrations
At Boster we strive to provide the best Anti-NM23A Antibody by testing all applications on non-spiked tissues and cell lines to ensure that the affinity of the antibody is enough to react to the endogenouse level of the target protein. Read more about our QC panel here.
|Recommended dilution ratios are listed below:|
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, By Heat|
Western blot, 0.1-0.5μg/ml, Human, Rat, Mouse
Immunohistochemistry(Frozen Section), 0.5-1μg/ml, Human
Immunocytochemistry, 0.5-1μg/ml, Human
Flow Cytometry, 1-3μg/1x106 cells, Human
**Boster provides high sensitivity secondary antibody kits for Western blotting and IHC. For more info see Related Products below.
Anti-NM23A Antibody Images
Click the images to enlarge.
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 Lysate
Lane 2: Rat Brain Tissue Lysate
Lane 3: Rat Liver Tissue Lysate
Lane 4: Rat Skeletal Muscle Tissue Lysate
Lane 5: PANC Cell Lysate
Lane 6: HELA Cell Lysate
Lane 7: SKOV Cell Lysate
Lane 8: COLO320 Cell Lysate
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- NM23A antigen affinity purified polyclonal antibody (Catalog # PA1829) 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 1:10000 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 NM23A at approximately 17KD. The expected band size for NM23A is at 17KD.
NM23A was detected in paraffin-embedded section of Rat Cerebellum 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- NM23A Antibody (PA1829) 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.
NM23A was detected in immunocytochemical section of HELA Cell. 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- NM23A Antibody (PA1829) 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.
Overlay histogram showing Hela cells stained with PA1829 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NME1 Antibody (PA1829,1μg/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.
1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,