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product-image-AAA19946_IF13.jpg IF (Immunofluorescence) (Figure 13. IF analysis of RNF25 using anti-RNF25 antibody (AAA19946).RNF25 was detected in a paraffin-embedded section of human lung cancer 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-RNF25 Antibody (AAA19946) overnight at 4 degree C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) 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.)

Rabbit RNF25 Polyclonal Antibody | anti-RNF25 antibody

Anti-RNF25 Antibody Picoband

Gene Names
RNF25; AO7
Reactivity
Human, Mouse, Rat
Applications
ELISA, Flow Cytometry, Functional Assay, Immunofluorescence, Immunohistochemistry, Western Blot
Purity
Immunogen affinity purified.
Synonyms
RNF25, Antibody; Anti-RNF25 Antibody Picoband; Serine/threonine-protein kinase AtPK1/AtPK6; 70 kDa ribosomal protein S6 kinase 1 antibody, KS6B1_HUMAN antibody, p70 alpha antibody, P70 beta 1 antibody, p70 ribosomal S6 kinase alpha antibody, p70 ribosomal S6 kinase beta 1 antibody, p70 S6 kinase alpha antibody, P70 S6 Kinase antibody, p70 S6 kinase alpha 1 antibody, p70 S6 kinase alpha 2 antibody, p70 S6K antibody, p70 S6K-alpha antibody, p70 S6KA antibody, p70(S6K) alpha antibody, p70(S6K)-alpha antibody, p70-alpha antibody, p70-S6K 1 antibody, p70-S6K antibody, P70S6K antibody, P70S6K1 antibody, p70S6Kb antibody, PS6K antibody, Ribosomal protein S6 kinase 70kDa polypeptide 1 antibody, Ribosomal protein S6 kinase beta 1 antibody, Ribosomal protein S6 kinase beta-1 antibody, Ribosomal protein S6 kinase I antibody, RPS6KB1 antibody, S6K antibody, S6K-beta-1 antibody, S6K1 antibody, Serine/threonine kinase 14 alpha antibody, Serine/threonine-protein kinase 14A antibody, STK14A antibody; anti-RNF25 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human, Mouse, Rat
Clonality
Polyclonal
Isotype
IgG
Specificity
Expressed in all 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-RNF25 antibody
ELISA (EIA), Flow Cytometry (FC/FACS), Immunofluorescence (IF), Immunohistochemistry (IHC), Western Blot (WB)
Application Notes
WB: 0.25-0.5ug/ml, Human, Mouse, Rat
IHC: 2-5ug/ml, Human, Rat
IF: 5ug/ml, Human
FC/FACS (Fixed): 1-3ug/1x106 cells, Human
ELISA: 0.1-0.5ug/ml
Immunogen
E coli-derived human RNF25 recombinant protein (Position: E44-R344). Human RNF25 shares 83.7% amino acid (aa) sequence identity with mouse RNF25.
Subcellular Localization
Cytoplasm, Nucleus
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
Downstream effector of TOR signaling pathway involved in osmotic stress response. Could be involved in the control of plant growth and development. Phosphorylates the ribosomal proteins P14, P16 and S6. Functions as a repressor of cell proliferation and required for maintenance of chromosome stability and ploidy levels through the RBR1-E2F pathway.
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.

IF (Immunofluorescence)

(Figure 13. IF analysis of RNF25 using anti-RNF25 antibody (AAA19946).RNF25 was detected in a paraffin-embedded section of human lung cancer 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-RNF25 Antibody (AAA19946) overnight at 4 degree C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) 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.)

product-image-AAA19946_IF13.jpg IF (Immunofluorescence) (Figure 13. IF analysis of RNF25 using anti-RNF25 antibody (AAA19946).RNF25 was detected in a paraffin-embedded section of human lung cancer 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-RNF25 Antibody (AAA19946) overnight at 4 degree C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) 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.)

FCM (Flow Cytometry)

(Figure 12. Flow Cytometry analysis of 293T cells using anti-RNF25 antibody (AAA19946).Overlay histogram showing 293T cells stained with AAA19946 (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-RNF25 Antibody (AAA19946, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10ug/1x106 cells) 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 (Red line) was also used as a control.)

product-image-AAA19946_FCM12.png FCM (Flow Cytometry) (Figure 12. Flow Cytometry analysis of 293T cells using anti-RNF25 antibody (AAA19946).Overlay histogram showing 293T cells stained with AAA19946 (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-RNF25 Antibody (AAA19946, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10ug/1x106 cells) 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 (Red line) was also used as a control.)

IHC (Immunohistochemistry)

(Figure 11. IHC analysis of RNF25 using anti-RNF25 antibody (AAA19946).RNF25 was detected in a paraffin-embedded section of rat testis 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 2ug/ml rabbit anti-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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 U251 whole cell lysates,Lane 3: human 293T whole cell lysates,Lane 4: human Jurkat whole cell lysates,Lane 5: rat testis tissue lysates,Lane 6: rat brain tissue lysates,Lane 7: mouse testis tissue lysates,Lane 8: mouse brain tissue 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-RNF25 antigen affinity purified polyclonal antibody (#AAA19946) 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 RNF25 at approximately 62 kDa. The expected band size for RNF25 is at 51 kDa.)

product-image-AAA19946_WB.jpg IHC (Immunohistochemistry) (Figure 11. IHC analysis of RNF25 using anti-RNF25 antibody (AAA19946).RNF25 was detected in a paraffin-embedded section of rat testis 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 2ug/ml rabbit anti-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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-RNF25 Antibody (AAA19946) 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 U251 whole cell lysates,Lane 3: human 293T whole cell lysates,Lane 4: human Jurkat whole cell lysates,Lane 5: rat testis tissue lysates,Lane 6: rat brain tissue lysates,Lane 7: mouse testis tissue lysates,Lane 8: mouse brain tissue 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-RNF25 antigen affinity purified polyclonal antibody (#AAA19946) 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 RNF25 at approximately 62 kDa. The expected band size for RNF25 is at 51 kDa.)
Related Product Information for anti-RNF25 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.
Product Categories/Family for anti-RNF25 antibody
References
1. Asamitsu, K., Tetsuka, T., Kanazawa, S., Okamoto, T. RING finger protein AO7 supports NF-kappa-B-mediated transcription by interacting with the transactivation domain of the p65 subunit. J. Biol. Chem. 278: 26879-26887, 2003.2. Hartz, P. A. Personal Communication. Baltimore, Md. 9/18/2014.3. Lorick, K. L., Jensen, J. P., Fang, S., Ong, A. M., Hatakeyama, S., Weissman, A. M. RING fingers mediate ubiquitin-conjugating enzyme (E2)-dependent ubiquitination. Proc. Nat. Acad. Sci. 96: 11364-11369, 1999.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
51,219 Da
NCBI Official Full Name
E3 ubiquitin-protein ligase RNF25
NCBI Official Synonym Full Names
ring finger protein 25
NCBI Official Symbol
RNF25
NCBI Official Synonym Symbols
AO7
NCBI Protein Information
E3 ubiquitin-protein ligase RNF25; ring finger protein AO7
UniProt Protein Name
E3 ubiquitin-protein ligase RNF25
UniProt Gene Name
RNF25
UniProt Entry Name
RNF25_HUMAN

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Product Notes

The RNF25 rnf25 (Catalog #AAA19946) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-RNF25 Antibody Picoband reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's RNF25 can be used in a range of immunoassay formats including, but not limited to, ELISA (EIA), Flow Cytometry (FC/FACS), Immunofluorescence (IF), Immunohistochemistry (IHC), Western Blot (WB). WB: 0.25-0.5ug/ml, Human, Mouse, Rat IHC: 2-5ug/ml, Human, Rat IF: 5ug/ml, Human FC/FACS (Fixed): 1-3ug/1x106 cells, Human ELISA: 0.1-0.5ug/ml. Researchers should empirically determine the suitability of the RNF25 rnf25 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 "RNF25, 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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