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Anti-Nav1.8/SCN10A Mouse Antibody [F24B8]

Catalog No.: F3462

Application: Reactivity: Mouse, Rat

Immunohistochemical analysis of formalin fixed paraffin embedded mouse skin tissue with F3462 at 1:100 dilution.

Usage Information

Dilution
IHC1:100

Application
IHC

Reactivity
Mouse, Rat

Source
Mouse

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Positive Control	Mouse dorsal root ganglia; Rat dorsal root ganglia
Negative Control

Exprimental Methods

IHC
Experimental Protocol： Deparaffinization/Rehydration 1. Deparaffinize/hydrate sections: 2. Incubate sections in three washes of xylene for 5 min each. 3. Incubate sections in two washes of 100% ethanol for 10 min each. 4. Incubate sections in two washes of 95% ethanol for 10 min each. 5. Wash sections two times in dH2O for 5 min each. 6.Antigen retrieval: For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; continue with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min. Staining 1. Wash sections in dH2O three times for 5 min each. 2. Incubate sections in 3% hydrogen peroxide for 10 min. 3. Wash sections in dH2O two times for 5 min each. 4. Wash sections in wash buffer for 5 min. 5. Block each section with 100–400 µl of blocking solution for 1 hr at room temperature. 6. Remove blocking solution and add 100–400 µl primary antibody diluent in to each section. Incubate overnight at 4°C. 7. Remove antibody solution and wash sections with wash buffer three times for 5 min each. 8. Cover section with 1–3 drops HRPas needed. Incubate in a humidified chamber for 30 min at room temperature. 9. Wash sections three times with wash buffer for 5 min each. 10. Add DAB Chromogen Concentrate to DAB Diluent and mix well before use. 11. Apply 100–400 µl DAB to each section and monitor closely. 1–10 min generally provides an acceptable staining intensity. 12. Immerse slides in dH2O. 13. If desired, counterstain sections with hematoxylin. 14. Wash sections in dH2O two times for 5 min each. 15. Dehydrate sections: Incubate sections in 95% ethanol two times for 10 sec each; Repeat in 100% ethanol, incubating sections two times for 10 sec each; Repeat in xylene, incubating sections two times for 10 sec each. 16. Mount sections with coverslips and mounting medium.

IHC

Experimental Protocol：

Deparaffinization/Rehydration

1. Deparaffinize/hydrate sections:

2. Incubate sections in three washes of xylene for 5 min each.

3. Incubate sections in two washes of 100% ethanol for 10 min each.

4. Incubate sections in two washes of 95% ethanol for 10 min each.

5. Wash sections two times in dH2O for 5 min each.

6.Antigen retrieval: For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; continue with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.

Staining

1. Wash sections in dH2O three times for 5 min each.

2. Incubate sections in 3% hydrogen peroxide for 10 min.

3. Wash sections in dH2O two times for 5 min each.

4. Wash sections in wash buffer for 5 min.

5. Block each section with 100–400 µl of blocking solution for 1 hr at room temperature.

6. Remove blocking solution and add 100–400 µl primary antibody diluent in to each section. Incubate overnight at 4°C.

7. Remove antibody solution and wash sections with wash buffer three times for 5 min each.

8. Cover section with 1–3 drops HRPas needed. Incubate in a humidified chamber for 30 min at room temperature.

9. Wash sections three times with wash buffer for 5 min each.

10. Add DAB Chromogen Concentrate to DAB Diluent and mix well before use.

11. Apply 100–400 µl DAB to each section and monitor closely. 1–10 min generally provides an acceptable staining intensity.

12. Immerse slides in dH2O.

13. If desired, counterstain sections with hematoxylin.

14. Wash sections in dH2O two times for 5 min each.

15. Dehydrate sections: Incubate sections in 95% ethanol two times for 10 sec each; Repeat in 100% ethanol, incubating sections two times for 10 sec each; Repeat in xylene, incubating sections two times for 10 sec each.

16. Mount sections with coverslips and mounting medium.

Datasheet & SDS

Biological Description

Specificity
Nav1.8/SCN10A Mouse mAb detects endogenous levels of total Nav1.8/SCN10A protein.

Subcellular Location
Cell membrane, Membrane

Uniprot ID
Q9Y5Y9

Clone
F24B8

Synonym(s)
Sodium channel protein type 10 subunit alpha, Peripheral nerve sodium channel 3, Sodium channel protein type X subunit alpha, Voltage-gated sodium channel subunit alpha Nav1.8, PN3, hPN3, SCN10A

Background
Nav1.8/SCN10A, encoded by the SCN10A gene, is a voltage-gated sodium channel α-subunit belonging to the TTX-resistant class, characterized by a cysteine substitution in its pore region that confers resistance to nanomolar tetrodotoxin inhibition. Structurally, it is a large transmembrane protein with four homologous domains (I–IV), each containing six membrane-spanning segments (S1–S6) that form the voltage-sensing and pore-forming regions, and is often associated with β-subunits that modulate gating and trafficking. NaV1.8 is predominantly expressed in small- and medium-diameter nociceptive sensory neurons of dorsal root ganglia (DRG) and cranial sensory ganglia, but is also present in peripheral neural elements of the heart, particularly intracardiac ganglia, while absent from cardiomyocytes. Functionally, NaV1.8 activates at relatively depolarized potentials, inactivates slowly, and maintains sodium currents at subthreshold voltages, enabling repetitive action potential firing and sustained excitability, especially at low temperatures. In DRG neurons, it plays a key role in pain signaling by amplifying excitability near the action potential threshold and interacting with NaV1.7, whereas in intracardiac neurons it regulates firing frequency and neurotransmitter release, potentially modulating cardiac conduction through neural control rather than direct myocyte depolarization.

Background

Nav1.8/SCN10A, encoded by the SCN10A gene, is a voltage-gated sodium channel α-subunit belonging to the TTX-resistant class, characterized by a cysteine substitution in its pore region that confers resistance to nanomolar tetrodotoxin inhibition. Structurally, it is a large transmembrane protein with four homologous domains (I–IV), each containing six membrane-spanning segments (S1–S6) that form the voltage-sensing and pore-forming regions, and is often associated with β-subunits that modulate gating and trafficking. NaV1.8 is predominantly expressed in small- and medium-diameter nociceptive sensory neurons of dorsal root ganglia (DRG) and cranial sensory ganglia, but is also present in peripheral neural elements of the heart, particularly intracardiac ganglia, while absent from cardiomyocytes. Functionally, NaV1.8 activates at relatively depolarized potentials, inactivates slowly, and maintains sodium currents at subthreshold voltages, enabling repetitive action potential firing and sustained excitability, especially at low temperatures. In DRG neurons, it plays a key role in pain signaling by amplifying excitability near the action potential threshold and interacting with NaV1.7, whereas in intracardiac neurons it regulates firing frequency and neurotransmitter release, potentially modulating cardiac conduction through neural control rather than direct myocyte depolarization.

References
https://pubmed.ncbi.nlm.nih.gov/39378238/ https://pubmed.ncbi.nlm.nih.gov/22723301/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%