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Purified anti-FOX3 (NeuN) Antibody (Previously Covance catalog# SIG-39860)

Pricing & Availability
Clone
1B7 (See other available formats)
Regulatory Status
RUO
Other Names
RNA binding protein fox-1 homolog 3, fox-1 homolog C, neuronal nuclei, hexaribonucleotide binding protein 3
Previously
Covance Catalog# SIG-39860
Isotype
Mouse IgG2b, κ
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Product Citations
publications
a-1B7_PURE_FOX3_NeuN_Antibody_IHC_1_010418
IHC staining of purified anti-FOX3 (NeuN) antibody (clone 1B7) on formalin-fixed paraffin-embedded mouse brain tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., the tissue was incubated with 0.5 µg/mL of the primary antibody overnight at 4°C. BioLegend’s Ultra Streptavidin (USA) HRP Detection Kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 4X objective. Scale bar: 300 µm
  • a-1B7_PURE_FOX3_NeuN_Antibody_IHC_1_010418
    IHC staining of purified anti-FOX3 (NeuN) antibody (clone 1B7) on formalin-fixed paraffin-embedded mouse brain tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., the tissue was incubated with 0.5 µg/mL of the primary antibody overnight at 4°C. BioLegend’s Ultra Streptavidin (USA) HRP Detection Kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 4X objective. Scale bar: 300 µm
  • c-1B7_PURE_FOX3_NeuN_Antibody_IHC_3_010418
    IHC staining of purified anti-FOX3 (NeuN) antibody (clone 1B7) on formalin-fixed paraffin-embedded mouse brain tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., the tissue was incubated with 0.5 µg/mL of the primary antibody overnight at 4°C. BioLegend’s Ultra Streptavidin (USA) HRP Detection Kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 10X objective. Scale bar: 100 µm
  • d-1B7_PURE_FOX3_NeuN_Antibody_IHC_4_010418
    IHC staining of purified anti-FOX3 (NeuN) antibody (clone 1B7) on formalin-fixed paraffin-embedded rat brain tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., the tissue was incubated with 0.5 µg/mL of the primary antibody overnight at 4°C. BioLegend’s Ultra Streptavidin (USA) HRP Detection Kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 10X objective. Scale bar: 100 µm
  • e-1B7_PURE_FOX3_NeuN_Antibody_IHC_WB_020520_updated
    Western blot of purified anti-FOX3 (NeuN) antibody (clone 1B7). Lane 1: Molecular weight marker; Lane 2: 20 µg of human brain lysate; Lane 3: 20 µg of mouse brain lysate; Lane 4: 20 µg of rat brain lysate. The blot was incubated with 0.05 µg/mL of the primary antibody overnight at 4°C, followed by incubation with HRP goat anti-mouse IgG antibody (Cat. No. 405306). Enhanced chemiluminescence was used as the detection system.
  • f-MCS-1B7_Purified_FOX3slashNeuN_Antibody_IHC_011615
    IHC staining of purified anti-FOX3 (NeuN) antibody (clone 1B7) on formalin-fixed paraffin-embedded human brain tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., the tissue was incubated with 1 µg/mL of the primary antibody for 60 minutes at room temperature. BioLegend’s Ultra Streptavidin (USA) HRP Detection Kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective.
  • g-1B7_PURE_FOX3_NeuN_Antibody_ICFC_7_040623
    A431 cells (low expression control, open histogram), or A549 cells (positive control, filled histogram) were fixed and permeabilized using True-Nuclear™ Transcription Factor Buffer Set (Cat. No. 424401) and intracellularly stained with purified anti-FOX3 (NeuN) (clone 1B7), or purified mouse IgG2b, κ isotype control (open histogram, dashed line) (representative histogram for both cell lines) (Cat. No. 400302), followed by PE goat anti-mouse IgG (Cat. No. 405307).
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834502 25 µL 112 CHF
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834501 100 µL 238 CHF
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Description

Fox3 is one of a family of 3 mammalian Fox homologues. Fox was discovered in C. elegans as a gene involved in sex determination, and the name Fox is an acronym of "Feminizing locus on X". The Fox protein and its 3 mammalian homologues are all about 46 kD proteins, each of which includes a central highly conserved RRM type RNA recognition motif, which corresponds to a small ~70 amino acid structure consisting of 4 beta strands and two alpha-helices. Fox3 is a protein which has a function in RNA splicing and is expressed heavily and specifically in neuronal nuclei. The 1B7 antibody reveals strong nuclear and distal cytoplasmic staining for Fox3/NeuN and the complete absence of staining of astrocytes and other kinds of non-neuronal cells. This Fox3/NeuN antibody is therefore an excellent marker of neuronal cells.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse, Rat
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Antibody was raised against the N-terminal 100 amino acids of human Fox3 as expressed in and purified from E. coli.
Formulation
Phosphate-buffered solution + 10mM sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
1 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C. Please note the storage condition for this antibody has been changed from -20°C to between 2°C and 8°C. You can also check your vial or your CoA to find the most accurate storage condition for this antibody.
Application

IHC-P -  Quality tested
WB, ICFC - Verified
ICC, IHC-F - Reported in the literature, not verified in house
SB - Community verified
Recommended Usage

Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry, a concentration range of 0.5 - 1.0 µg/mL is suggested. For western blotting, the suggested use of this reagent is 0.01 - 0.05 µg/mL. For intracellular flow cytometric staining, the suggested use of this reagent is ≤ 0.125 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

The double band observed in the immunoblot corresponds to the two NeuN isoforms produced by alternative splicing with expected molecular weights of 46 kD and 48 kD.

Additional Product Notes

This product has been verified for IHC-F (Immunohistochemistry - frozen tissue sections) on the NanoString GeoMx® Digital Spatial Profiler. The GeoMx® enables researchers to perform spatial analysis of protein and RNA targets in FFPE and fresh frozen human and mouse samples. For more information about our spatial biology products and the GeoMx® platform, please visit our spatial biology page.

Application References

(PubMed link indicates BioLegend citation)
  1. Herculano-Houzel S and Lent R. 2005. J Neurosci. 25:2518-21. (ICC)
  2. Kim KK, et al. 2009. J Biol Chem. 284:31052-31061. (WB, ICC, IHC-P)
  3. Underwood JG, et al. 2005. Mol Cell Biol. 25:10005-10016. (IHC-F, WB)
Product Citations
  1. Sun Y, et al. 2020. J Neuroinflammation. 17:72. PubMed
  2. Wang S, et al. 2022. PLoS Pathog. 18:e1010281. PubMed
  3. Dhekne HS, et al. 2018. Elife. 7:e40202. PubMed
  4. Yasom S, et al. 2022. FASEB Bioadv. 4:408. PubMed
  5. Schwabenland M, et al. 2021. Immunity. . PubMed
  6. He Y, et al. 2021. Sci Rep. 11:21348. PubMed
  7. Rathnam C, et al. 2021. Sci Adv. 7:eabj2281. PubMed
RRID
AB_2734601 (BioLegend Cat. No. 834502)
AB_2734601 (BioLegend Cat. No. 834501)

Antigen Details

Structure
Expected MW: 46 kD and 48 kD
Cell Type
Mature Neurons, Neural Stem Cells
Biology Area
Cell Biology, Neuroscience, Neuroscience Cell Markers, Stem Cells
Molecular Family
Nuclear Markers
Gene ID
146713 View all products for this Gene ID
UniProt
View information about FOX3 on UniProt.org

Related FAQs

If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?

It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.

Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?

Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.

Are other fluorophores compatible with IBEX?

Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.

The same antibody works in one tissue type but not another. What is happening?

Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.

How can I be sure the staining I’m seeing in my tissue is real?

In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.

Other Formats

View All FOX3 Reagents Request Custom Conjugation
Description Clone Applications
Purified anti-FOX3 (NeuN) 1B7 IHC-P,WB,ICFC,ICC,IHC-F,SB
Go To Top Version: 7    Revision Date: 01.24.2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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