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Mature Neuron Marker Antibody Kit

RMB: 5300

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Catalog# HAK21007

Mature Neuron Marker Antibody Kit

Application

  • WB

Reactivity

  • Human

  • Mouse

  • Rat

General Overview

Kit Components

Product Includes Specification Application Reactivity Mw
NeuN[ET1602-12] 20µl WB,IF-Cell,IF-Tissue,IHC-P,FC,IHC-Fr,mIHC Human,Mouse,Rat Predicted band size: 34 kDa
GAP43[ET1610-94] 20µl WB,IF-Cell,IF-Tissue,IHC-P,FC,IP Human,Mouse,Rat Predicted band size: 25 kDa
MAP2[HA723025] 20µl WB,IHC-P,IHC-Fr,IF-Cell Human,Mouse,Rat Predicted band size: 200 kDa
NF-L[HA721538] 20µl WB,IHC-P,IF-Cell,IF-Tissue,IHC-Fr Human,Mouse,Rat Predicted band size: 62 kDa
Beta III Tubulin[ET1604-17] 20µl WB,IHC-P,IP,FC,IHC-Fr,IF-Cell Human,Mouse,Rat Predicted band size: 50 kDa
Synaptophysin[ET1606-56] 20µl WB,IHC-P,IHC-Fr,IF-Tissue Human,Mouse,Rat Predicted band size: 34 kDa
PSD95[ET1602-20] 20µl WB,IF-Tissue,IHC-P,IHC-Fr Human,Mouse,Rat Predicted band size: 80 kDa
PGP9.5[ET1703-22] 20µl WB,IF-Cell,IF-Tissue,IHC-P,IP,FC,IHC-Fr Human,Mouse,Rat Predicted band size: 25 kDa
Alpaca anti-Rabbit IgG Fc, Recombinant VHH[HA1031] 100µl IP,ELISA,IHC-P,WB Rabbit

Product Description

Neuronal nuclei (NeuN, Fox-3, RBFOX3) is a nuclear protein expressed in most post-mitotic neurons of the central and peripheral nervous systems. NeuN is not detected in Purkinje cells, sympathetic ganglion cells, Cajal-Retzius cells, INL retinal cells, inferior olivary, and dentate nucleus neurons. This neuronal protein was originally identified by immunoreactivity with a monoclonal antibody also called NeuN. Using MS-analysis, NeuN was later identified as the Fox-3 gene product. Fox-3 contains an RNA recognition motif and functions as a splicing regulator. Fox-3 regulates alternative splicing of NumB, promoting neuronal differentiation during development.

Product Features

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Background

The antibodies in this kit serve to characterize and identify mature neurons. Neuronal nuclei (NeuN, Fox-3, RBFOX3) is a nuclear protein expressed in most post-mitotic neurons of the central and peripheral nervous systems. </br>GAP43 is a nervous system specific, growth-associated protein enriched in growth cones and areas of high plasticity. β3-tubulin is one of six β-tubulin isoforms that make up the building blocks of microtubules. Microtubule-associated protein 2 (MAP2) is a neuronal phosphoprotein that regulates the structure and stability of microtubules, neuronal morphogenesis, cytoskeleton dynamics, and organelle trafficking in axons and dendrites. Neurofilaments are the major intermediate filaments found in neurons and consist of light (NFL), medium (NFM), and heavy (NFH) subunits. Synaptophysin is a neuronal synaptic vesicle glycoprotein. Synaptophysin is responsible for targeting synaptobrevin 2/VAMP2 to synaptic vesicles, and is a critical component and marker for the presynaptic fusion complex. PSD95 is a scaffolding protein involved in the assembly and function of mature postsynaptic density complexes.

Data Links

Background References

1. Mullen, R.J. et al. (1992) Development 116, 201-11.

2. Kim, K.K. et al. (2009) J Biol Chem 284, 31052-61.

3. Biewenga, J.E. et al. (1996) Acta Biochim Pol 43, 327-38.

4. Aigner, L. and Caroni, P. (1993) J Cell Biol 123, 417-29.

5. Jiang, Y.Q. and Oblinger, M.M. (1992) J Cell Sci 103 (Pt 3), 643-51.

6. Sánchez, C. et al. (2000) Prog Neurobiol 61, 133-68.

7. Caceres, A. et al. (1984) Brain Res 315, 314-8.

8. Al-Chalabi, A. and Miller, C.C. (2003) Bioessays 25, 346-55.

Images

  • Western blot analysis of NeuN on different lysates with Rabbit anti-NeuN antibody (<a href="/products/ET1602-12" style="font-weight: bold;text-decoration: underline;">ET1602-12</a>) at 1/5,000 dilution.<br /><br />Lane 1: Mouse brain tissue lysate (20 µg/Lane)<br />Lane 2: Rat brain tissue lysate (20 µg/Lane)<br />Lane 3: Mouse cerebellum tissue lysate (20 µg/Lane)<br />Lane 4: Rat cerebellum tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 34 kDa<br />Observed band size: 45/50 kDa<br /><br />Exposure time: 43 seconds;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1602-12" style="font-weight: bold;text-decoration: underline;">ET1602-12</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of NeuN on different lysates with Rabbit anti-NeuN antibody (ET1602-12) at 1/5,000 dilution.

    Lane 1: Mouse brain tissue lysate (20 µg/Lane)
    Lane 2: Rat brain tissue lysate (20 µg/Lane)
    Lane 3: Mouse cerebellum tissue lysate (20 µg/Lane)
    Lane 4: Rat cerebellum tissue lysate (20 µg/Lane)

    Predicted band size: 34 kDa
    Observed band size: 45/50 kDa

    Exposure time: 43 seconds;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1602-12) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of GAP43 on different lysates with Rabbit anti-GAP43 antibody (<a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>) at 1/5,000 dilution.<br /><br />Lane 1: SH-SY5Y cell lysate (20 µg/Lane)<br />Lane 2: Neuro-2a cell lysate (20 µg/Lane)<br />Lane 3: Mouse brain tissue lysate (20 µg/Lane)<br />Lane 4: Rat brain tissue lysate (20 µg/Lane)<br />Lane 5: Rat hippocampus tissue lysate (20 µg/Lane)<br />Lane 6: Mouse hippocampus tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 25 kDa<br />Observed band size: 43/45 kDa<br /><br />Exposure time: 24 seconds;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of GAP43 on different lysates with Rabbit anti-GAP43 antibody (ET1610-94) at 1/5,000 dilution.

    Lane 1: SH-SY5Y cell lysate (20 µg/Lane)
    Lane 2: Neuro-2a cell lysate (20 µg/Lane)
    Lane 3: Mouse brain tissue lysate (20 µg/Lane)
    Lane 4: Rat brain tissue lysate (20 µg/Lane)
    Lane 5: Rat hippocampus tissue lysate (20 µg/Lane)
    Lane 6: Mouse hippocampus tissue lysate (20 µg/Lane)

    Predicted band size: 25 kDa
    Observed band size: 43/45 kDa

    Exposure time: 24 seconds;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-94) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of GAP43 on different lysates with Rabbit anti-GAP43 antibody (<a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>) at 1/100,000 dilution and competitor's antibody at 1/100,000 dilution.<br /><br />Lane 1: SH-SY5Y cell lysate (20 µg/Lane)<br />Lane 2: A549 cell lysate (negative) (20 µg/Lane)<br />Lane 3: Neuro-2a cell lysate (20 µg/Lane)<br />Lane 4: Mouse lung tissue lysate (negative) (20 µg/Lane)<br />Lane 5: Mouse brain tissue lysate (20 µg/Lane)<br />Lane 6: Rat brain tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 25 kDa<br />Observed band size: 43/45 kDa<br /><br />Exposure time: 3 minutes 20 seconds;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>) at 1/100,000 dilution and competitor's antibody at 1/100,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of GAP43 on different lysates with Rabbit anti-GAP43 antibody (ET1610-94) at 1/100,000 dilution and competitor's antibody at 1/100,000 dilution.

    Lane 1: SH-SY5Y cell lysate (20 µg/Lane)
    Lane 2: A549 cell lysate (negative) (20 µg/Lane)
    Lane 3: Neuro-2a cell lysate (20 µg/Lane)
    Lane 4: Mouse lung tissue lysate (negative) (20 µg/Lane)
    Lane 5: Mouse brain tissue lysate (20 µg/Lane)
    Lane 6: Rat brain tissue lysate (20 µg/Lane)

    Predicted band size: 25 kDa
    Observed band size: 43/45 kDa

    Exposure time: 3 minutes 20 seconds;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-94) at 1/100,000 dilution and competitor's antibody at 1/100,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of Beta III Tubulin on different lysates with Rabbit anti-Beta III Tubulin antibody (<a href="/products/ET1604-17" style="font-weight: bold;text-decoration: underline;">ET1604-17</a>) at 1/20,000 dilution.<br /><br />Lane 1: SH-SY5Y cell lysate (15 µg/Lane)<br />Lane 2: HeLa cell lysate (15 µg/Lane)<br />Lane 3: Neuro-2a cell lysate (15 µg/Lane)<br />Lane 4: NIH/3T3 cell lysate (15 µg/Lane)<br />Lane 5: PC-12 cell lysate (15 µg/Lane)<br />Lane 6: C6 cell lysate (15 µg/Lane)<br />Lane 7: Mouse brain tissue lysate (20 µg/Lane)<br />Lane 8: Rat brain tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 50 kDa<br />Observed band size: 50 kDa<br /><br />Exposure time: 3 minutes 54 seconds;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1604-17" style="font-weight: bold;text-decoration: underline;">ET1604-17</a>) at 1/20,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Beta III Tubulin on different lysates with Rabbit anti-Beta III Tubulin antibody (ET1604-17) at 1/20,000 dilution.

    Lane 1: SH-SY5Y cell lysate (15 µg/Lane)
    Lane 2: HeLa cell lysate (15 µg/Lane)
    Lane 3: Neuro-2a cell lysate (15 µg/Lane)
    Lane 4: NIH/3T3 cell lysate (15 µg/Lane)
    Lane 5: PC-12 cell lysate (15 µg/Lane)
    Lane 6: C6 cell lysate (15 µg/Lane)
    Lane 7: Mouse brain tissue lysate (20 µg/Lane)
    Lane 8: Rat brain tissue lysate (20 µg/Lane)

    Predicted band size: 50 kDa
    Observed band size: 50 kDa

    Exposure time: 3 minutes 54 seconds;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1604-17) at 1/20,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of Synaptophysin on different lysates with Rabbit anti-Synaptophysin antibody (<a href="/products/ET1606-56" style="font-weight: bold;text-decoration: underline;">ET1606-56</a>) at 1/5,000 dilution.<br /><br />Lane 1: SH-SY5Y cell lysate (20 µg/Lane)<br />Lane 2: HeLa cell lysate (negative) (20 µg/Lane)<br />Lane 3: NIH/3T3 cell lysate (negative) (20 µg/Lane)<br />Lane 4: PC-12 cell lysate (20 µg/Lane)<br />Lane 5: Human brain tissue lysate (20 µg/Lane)<br />Lane 6: Mouse brain tissue lysate (20 µg/Lane)<br />Lane 7: Rat brain tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 34 kDa<br />Observed band size: 34/40 kDa<br /><br />Exposure time: 43 seconds;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1606-56" style="font-weight: bold;text-decoration: underline;">ET1606-56</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Synaptophysin on different lysates with Rabbit anti-Synaptophysin antibody (ET1606-56) at 1/5,000 dilution.

    Lane 1: SH-SY5Y cell lysate (20 µg/Lane)
    Lane 2: HeLa cell lysate (negative) (20 µg/Lane)
    Lane 3: NIH/3T3 cell lysate (negative) (20 µg/Lane)
    Lane 4: PC-12 cell lysate (20 µg/Lane)
    Lane 5: Human brain tissue lysate (20 µg/Lane)
    Lane 6: Mouse brain tissue lysate (20 µg/Lane)
    Lane 7: Rat brain tissue lysate (20 µg/Lane)

    Predicted band size: 34 kDa
    Observed band size: 34/40 kDa

    Exposure time: 43 seconds;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1606-56) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of PSD95 on different lysates with Rabbit anti-PSD95 antibody (<a href="/products/ET1602-20" style="font-weight: bold;text-decoration: underline;">ET1602-20</a>) at 1/2,000 dilution.<br /><br />Lane 1: C6 cell lysate (15 µg/Lane)<br />Lane 2: Mouse brain tissue lysate (20 µg/Lane)<br />Lane 3: Rat brain tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 80 kDa<br />Observed band size: 100 kDa<br /><br />Exposure time: 5 minutes 10 seconds;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1602-20" style="font-weight: bold;text-decoration: underline;">ET1602-20</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of PSD95 on different lysates with Rabbit anti-PSD95 antibody (ET1602-20) at 1/2,000 dilution.

    Lane 1: C6 cell lysate (15 µg/Lane)
    Lane 2: Mouse brain tissue lysate (20 µg/Lane)
    Lane 3: Rat brain tissue lysate (20 µg/Lane)

    Predicted band size: 80 kDa
    Observed band size: 100 kDa

    Exposure time: 5 minutes 10 seconds;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1602-20) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

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