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FCM (Flow Cytometry) (Figure 6. Flow Cytometry analysis of RT4 cells using anti-RFX3 antibody (AAA19892).Overlay histogram showing RT4 cells stained with AAA19892 (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-RFX3 Antibody (AAA19892, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.)

Rabbit RFX3 Polyclonal Antibody | anti-RFX3 antibody

Anti-RFX3 Antibody Picoband

Reactivity
Human, Mouse, Rat
Applications
Western Blot, Immunohistochemistry, Immunocytochemistry, Immunofluorescence, Flow Cytometry, Functional Assay, ELISA
Purity
Immunogen affinity purified.
Synonyms
RFX3; Polyclonal Antibody; Anti-RFX3 Antibody Picoband; BAG family molecular chaperone regulator 5; cadherin 4; Cadherin-4; Retinal cadherin; R-CAD; R-cadherin; CDH4; anti-RFX3 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human, Mouse, Rat
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Expressed mainly in brain but also found in other tissues.
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized. Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Applicable Applications for anti-RFX3 antibody
Western Blot (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS), ELISA (EIA)
Application Notes
WB: 0.25-0.5ug/ml, Human
IHC(Paraffin-embedded Section): 2-5ug/ml, Mouse, Rat
ICC/IF: 5ug/ml, Human
IF: 5ug/ml, Rat
FC/FACS (Fixed): 1-3ug/1x106 cells, Human
ELISA: 0.1-0.5ug/ml
Immunogen
E coli-derived human RFX3 recombinant protein (Position: Q283-V749).
Subcellular Localization
Cell membrane. Single-pass type I membrane protein
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500ug/ml.
Cross Reactivity
No cross-reactivity with other proteins.
Protein Function
Cadherins are calcium-dependent cell adhesion proteins. They preferentially interact with themselves in a homophilic manner in connecting cells; cadherins may thus contribute to the sorting of heterogeneous cell types. May play an important role in retinal development.
Preparation and Storage
At -20 degree C for one year from date of receipt. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for six months.Avoid repeated freezing and thawing.

FCM (Flow Cytometry)

(Figure 6. Flow Cytometry analysis of RT4 cells using anti-RFX3 antibody (AAA19892).Overlay histogram showing RT4 cells stained with AAA19892 (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-RFX3 Antibody (AAA19892, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.)

FCM (Flow Cytometry) (Figure 6. Flow Cytometry analysis of RT4 cells using anti-RFX3 antibody (AAA19892).Overlay histogram showing RT4 cells stained with AAA19892 (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-RFX3 Antibody (AAA19892, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.)

IF (Immunofluorescence)

(Figure 5. IF analysis of RFX3 using anti-RFX3 antibody (AAA19892).RFX3 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 5ug/mL rabbit anti-RFX3 Antibody (AAA19892) overnight at 4 degree C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.)

IF (Immunofluorescence) (Figure 5. IF analysis of RFX3 using anti-RFX3 antibody (AAA19892).RFX3 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 5ug/mL rabbit anti-RFX3 Antibody (AAA19892) overnight at 4 degree C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.)

IF (Immunofluorescence)

(Figure 4. IF analysis of RFX3 using anti-RFX3 antibody (AAA19892) and anti-Tubulin Alpha antibody (M03989-3).RFX3 was detected in immunocytochemical section of U2OS cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-RFX3 Antibody (AAA19892) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-RFX3 Antibody (AAA19892) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Lane 2: human SH-SY5Y 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-RFX3 antigen affinity purified polyclonal antibody (#AAA19892) at 0.5ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for RFX3 at approximately 95 kDa. The expected band size for RFX3 is at 84 kDa.)

IF (Immunofluorescence) (Figure 4. IF analysis of RFX3 using anti-RFX3 antibody (AAA19892) and anti-Tubulin Alpha antibody (M03989-3).RFX3 was detected in immunocytochemical section of U2OS cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-RFX3 Antibody (AAA19892) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-RFX3 Antibody (AAA19892) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Lane 2: human SH-SY5Y 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-RFX3 antigen affinity purified polyclonal antibody (#AAA19892) at 0.5ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for RFX3 at approximately 95 kDa. The expected band size for RFX3 is at 84 kDa.)
Related Product Information for anti-RFX3 antibody
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.
References
1. Baas, D., Meiniel, A., Benadiba, C., Bonnafe, E., Meiniel, O., Reith, W., Durand, B. A deficiency in RFX3 causes hydrocephalus associated with abnormal differentiation of ependymal cells. Europ. J. Neurosci. 24: 1020-1030, 2006. 2. Bonnafe, E., Touka, M., AitLounis, A., Baas, D., Barras, E., Ucla, C., Moreau, A., Flamant, F., Dubruille, R., Couble, P., Collignon, J., Durand, B., Reith, W. The transcription factor RFX3 s nodal cilium development and left-right asymmetry specification. Molec. Cell. Biol. 24: 4417-4427, 2004.3. Elkon, R., Milon, B., Morrison, L., Shah, M., Vijayakumar, S., Racherla, M., Leitch, C. C., Silipino, L., Hadi, S., Weiss-Gayet, M., Barras, E., Schmid, C. D., and 12 others. RFX transcription factors are essential for hearing in mice. Nature Commun. 6: 8549, 2015. Note: Electronic Article.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
45,715 Da
NCBI Official Full Name
transcription factor RFX3 isoform a
NCBI Official Synonym Full Names
regulatory factor X, 3 (influences HLA class II expression)
NCBI Official Symbol
RFX3
NCBI Protein Information
transcription factor RFX3; DNA binding protein RFX3
UniProt Protein Name
Transcription factor RFX3
UniProt Gene Name
RFX3
UniProt Entry Name
RFX3_HUMAN

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Product Notes

The RFX3 rfx3 (Catalog #AAA19892) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-RFX3 Antibody Picoband reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's RFX3 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS), ELISA (EIA). WB: 0.25-0.5ug/ml, Human IHC(Paraffin-embedded Section): 2-5ug/ml, Mouse, Rat ICC/IF: 5ug/ml, Human IF: 5ug/ml, Rat FC/FACS (Fixed): 1-3ug/1x106 cells, Human ELISA: 0.1-0.5ug/ml. Researchers should empirically determine the suitability of the RFX3 rfx3 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "RFX3, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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