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Product Info Summary
| SKU: | PA1830 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-NME2 Antibody Picoband®
SKU/Catalog Number
PA1830
BA3788 is an alternative SKU for this antibody, used in previous lots.
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. 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.
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 Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1830)
Host
Rabbit
Contents
Each vial contains antibody formulated with stabilizing components, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.
*This antibody is supplied in a stabilized formulation.
Compatibility with conjugation reactions depends on the chemistry of the conjugation method used.
For conjugation methods that are not compatible with the stabilizing components present in this formulation, a carrier-free antibody format is required.
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.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PA1830 is reactive to NME2 in Human, Mouse, Rat
Observed Molecular Weight
17 kDa
Calculated molecular weight
17.3 kDa
Background of 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.
Application & Images
Applications
PA1830 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
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
Immunocytochemistry , 0.5-1μg/ml, Human
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: rat heart tissue, rat brain tissue, mouse heart tissue, mouse brain tissue,
IHC: Human Intestinal Cancer Tissue, Rat Cerebellum Tissue, mouse intestine tissue
ICC/IF: U20S cell
ICC: HELA Cell
FCM: HL-60 cell
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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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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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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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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Flow Cytometry analysis of HL-60 cells using anti-NME2 antibody (PA1830).
Overlay histogram showing HL-60 cells stained with PA1830 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-NME2 Antibody Picoband® (PA1830)
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4 Customer Q&As for Anti-NME2 Antibody Picoband®
Question
Does anti-NME2 antibody PA1830 work for IHC-P with putamen?
Verified Customer
Verified customer
Asked: 2019-05-30
Answer
According to the expression profile of putamen, NME2 is highly expressed in putamen. So, it is likely that anti-NME2 antibody PA1830 will work for IHC-P with putamen.
Boster Scientific Support
Answered: 2019-05-30
Question
We are currently using anti-NME2 antibody PA1830 for rat tissue, and we are satisfied with the WB results. The species of reactivity given in the datasheet says human, mouse, rat. Is it true that the antibody can work on monkey tissues as well?
Verified Customer
Verified customer
Asked: 2019-03-18
Answer
The anti-NME2 antibody (PA1830) has not been validated for cross reactivity specifically with monkey tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in monkey you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-03-18
Question
I see that the anti-NME2 antibody PA1830 works with IHC-P, what is the protocol used to produce the result images on the product page?
M. Wu
Verified customer
Asked: 2015-11-11
Answer
You can find protocols for IHC-P on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to support@bosterbio.com
Boster Scientific Support
Answered: 2015-11-11
Question
I was wanting to use your anti-NME2 antibody for IHC-P for human putamen on frozen tissues, but I want to know if it has been tested for this particular application. Has this antibody been tested and is this antibody a good choice for human putamen identification?
P. Kulkarni
Verified customer
Asked: 2013-12-04
Answer
You can see on the product datasheet, PA1830 anti-NME2 antibody has been tested for IHC-P, ICC, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in human putamen in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2013-12-04
