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Product Info Summary
| SKU: | A14597-3 |
|---|---|
| Size: | 100 µg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-NXPH3 Antibody Picoband®
SKU/Catalog Number
A14597-3
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-NXPH3 Antibody Picoband® catalog # A14597-3. Tested in WB, IHC, ICC/IF, FCM, ELISA 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
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 freezing and thawing.
Cite This Product
Anti-NXPH3 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A14597-3)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Immunogen
E.coli-derived human NXPH3 recombinant protein (Position: R5-K217). Human NXPH3 shares 95.8% amino acid (aa) sequence identity with mouse and rat NXPH3, respectively.
Reactive Species
A14597-3 is reactive to NXPH3 in Human, Mouse, Rat
Observed Molecular Weight
28 kDa
Calculated molecular weight
28.1 kDa
Background of NXPH3
Neurexophilin-3 is a protein that in humans is encoded by the NXPH3 gene. Neurexophilin-3 (NXPH-3) is a neuropeptide-like secreted glycoprotein in the neurexophilin family. The 252 amino acid (aa) NXPH-3 precursor contains a 22 aa signal peptide, plus a 230 aa proprecursor that is likely cleaved at a basic motif, producing a 76 aa propeptide and a 154 aa mature protein. Mature rat NXPH-3 shares 99%, 95% and 92% aa identity with mouse, human and bovine NXPH-3, respectively. It contains motifs that are conserved among all neurexophilins, including three potential N-glycosylation sites in the N-terminal portion and six cysteines in the C-terminal portion. NXPH-3 is expressed selectively in subplate-derived neurons in the cortex, granule cells in the vestibulocerebellum, and Cajal-Retzius cells during development. NXPH-1 is the neurexophilin most similar to NXPH-3, sharing 69% aa identity within the mature region. Expression of NXPH-1 and NXPH-3 does not appear to overlap, with NXPH-1 expression occurring mainly by cells that resemble inhibitory interneurons. Both are tightly bound extracellular ligands of alpha -neurexins, synaptic transmembrane molecules that are essential for calcium-triggered neurotransmitter release. Genetic deletion of NXPH-1 and/or NXPH-3 produces no anatomical effect, although mice lacking NXPH-3 show specific defects in motor coordination. Of the other known neurexophilins, NXPH-2 is not expressed in rodents, and NXPH-4 does not bind alpha -neurexins.
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
A14597-3 is guaranteed for ELISA, 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, 0.25-0.5 μg/ml, Human
Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human 293T whole cell, human MCF-7 whole cell, human HepG2 whole cell
IHC: mouse brain tissue, mouse brain tissue, rat brain tissue, rat brain tissue
ICC/IF: U2OS cell
FCM: HepG2 cell, K562 cell
Validation Images & Assay Conditions
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Western blot analysis of NXPD3 using anti-NXPD3 antibody (A14597-3).
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 30 ug of sample under reducing conditions.
Lane 1: human 293T whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: human HepG2 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-NXPD3 antigen affinity purified polyclonal antibody (Catalog # A14597-3) 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 NXPD3 at approximately 28 kDa. The expected band size for NXPD3 is at 28 kDa.
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IHC analysis of Neurexophilin 3/NXPH3 using anti-Neurexophilin 3/NXPH3 antibody (A14597-3).
Neurexophilin 3/NXPH3 was detected in a paraffin-embedded section of human brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Neurexophilin 3/NXPH3 Antibody (A14597-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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IHC analysis of NXPD3 using anti-NXPD3 antibody (A14597-3).
NXPD3 was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NXPD3 Antibody (A14597-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
IHC analysis of NXPD3 using anti-NXPD3 antibody (A14597-3).
NXPD3 was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NXPD3 Antibody (A14597-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
IHC analysis of NXPD3 using anti-NXPD3 antibody (A14597-3).
NXPD3 was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NXPD3 Antibody (A14597-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
IHC analysis of NXPD3 using anti-NXPD3 antibody (A14597-3).
NXPD3 was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NXPD3 Antibody (A14597-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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IF analysis of NXPD3 using anti-NXPD3 antibody (A14597-3) and anti-Beta Tubulin antibody (M01857-3).
NXPD3 was detected in immunocytochemical section of U2OS cell. 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 5 μg/mL rabbit anti-NXPD3 Antibody (A14597-3) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) and DyLight®550 Conjugated Goat Anti-Mouse IgG (BA1133) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Flow Cytometry analysis of HepG2 cells using anti-NXPH3 antibody (A14597-3).
Overlay histogram showing HepG2 cells stained with A14597-3 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-NXPH3 Antibody (A14597-3, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
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Flow Cytometry analysis of K562 cells using anti-NXPH3 antibody (A14597-3).
Overlay histogram showing K562 cells stained with A14597-3 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-NXPH3 Antibody (A14597-3, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Specific Publications For Anti-NXPH3 Antibody Picoband® (A14597-3)
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