Western Blot (WB)
(Figure 1. Western blot analysis of RNF8 using anti-RNF8 antibody (MBS1753365).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: rai brain tissue lysatesLane 2: mouse brain tissue lysatesLane 3: mouse NIH/3T3 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-RNF8 antigen affinity purified polyclonal antibody (Catalog # MBS1753365) at 0. 25 μg/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 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for RNF8 at approximately 56KD. The expected band size for RNF8 is at 56KD. )
Rabbit anti-Mouse, Rat RNF8 Polyclonal Antibody | anti-Rnf8 antibody
Anti-RNF8 Antibody
Gene Names
Rnf8; AIP37; 3830404E21Rik
Reactivity
Mouse, Rat
Applications
Western Blot, Flow Cytometry, Functional Assay, ELISA
Purity
Immunogen affinity purified.
Synonyms
RNF8; Polyclonal Antibody; Anti-RNF8 Antibody; Rnf8; E3 ubiquitin-protein ligase RNF8; EC 2. 3. 2. 27; ActA-interacting protein 37; AIP37; LaXp180; RING finger protein 8; RING-type E3 ubiquitin transferase RNF8; ring finger protein 8; anti-Rnf8 antibody
Host
Rabbit
Reactivity
Mouse, Rat
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Rabbit IgG polyclonal antibody for RNF8 detection.
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized. Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.
Applicable Applications for anti-Rnf8 antibody
Western Blot (WB), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA)
Application Notes
WB: 0.1-0.25ug/ml|Mouse, Rat|
FC/FACS/FCM: 1-3ug/1x106 cells|Mouse|
Direct ELISA: 0.1-0.5ug/ml|Mouse|
FC/FACS/FCM: 1-3ug/1x106 cells|Mouse|
Direct ELISA: 0.1-0.5ug/ml|Mouse|
Immunogen
E Coli-derived mouse RNF8 recombinant protein (Position: E3-S488).
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Recommended Detection Systems
Recommended Detection Systems
Preparation and Storage
Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.
Western Blot (WB)
(Figure 1. Western blot analysis of RNF8 using anti-RNF8 antibody (MBS1753365).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: rai brain tissue lysatesLane 2: mouse brain tissue lysatesLane 3: mouse NIH/3T3 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-RNF8 antigen affinity purified polyclonal antibody (Catalog # MBS1753365) at 0. 25 μg/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 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for RNF8 at approximately 56KD. The expected band size for RNF8 is at 56KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of RNF8 using anti-RNF8 antibody (MBS1753365).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: rai brain tissue lysatesLane 2: mouse brain tissue lysatesLane 3: mouse NIH/3T3 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-RNF8 antigen affinity purified polyclonal antibody (Catalog # MBS1753365) at 0. 25 μg/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 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for RNF8 at approximately 56KD. The expected band size for RNF8 is at 56KD. )
Flow Cytometry (FC/FACS)
(Figure 2. Flow Cytometry analysis of RAW264. 7 cells using anti-RNF8 antibody (MBS1753365).Overlay histogram showing RAW264. 7 cells stained with MBS1753365 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-RNF8 Antibody (MBS1753365,1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 2. Flow Cytometry analysis of RAW264. 7 cells using anti-RNF8 antibody (MBS1753365).Overlay histogram showing RAW264. 7 cells stained with MBS1753365 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-RNF8 Antibody (MBS1753365,1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Related Product Information for anti-Rnf8 antibody
E3 ubiquitin-protein ligase RNF8 is an enzyme that in humans is encoded by the RNF8 gene. The protein encoded by this gene contains a RING finger motif and an FHA domain. This protein has been shown to interact with several class II ubiquitin-conjugating enzymes (E2), including UBE2E1/UBCH6, UBE2E2, and UBE2E3, and may act as an ubiquitin ligase (E3) in the ubiquitination of certain nuclear proteins. This protein is also known to play a role in the DNA damage response and depletion of this protein causes cell growth inhibition and cell cycle arrest. Alternative splicing results in multiple transcript variants.
References
1. Bekker-Jensen, S., Rendtlew Danielsen, J., Fugger, K., Gromova, I., Nerstedt, A., Lukas, C., Bartek, J., Lukas, J., Mailand, N. HERC2 coordinates ubiquitin-dependent assembly of DNA repair factors on damaged chromosomes. Nature Cell Biol. 12: 80-86, 2010. Note: Erratum Nature Cell Biol. 12: 412 only, 2010.
2. Ishikawa, K., Nagase, T., Suyama, M., Miyajima, N., Tanaka, A., Kotani, H., Nomura, N., Ohara, O. Prediction of the coding sequences of unidentified human genes. X. The complete sequences of 100 new cDNA clones from brain which can code for large proteins in vitro. DNA Res. 5: 169-176, 1998.
3. Kolas, N. K., Chapman, J. R., Nakada, S., Ylanko, J., Chahwan, R., Sweeney, F. D., Panier, S., Mendez, M., Wildenhain, J., Thomson, T. M., Pelletier, L., Jackson, S. P., Durocher, D. Orchestration of the DNA-damage response by the RNF8 ubiquitin ligase. Science 318: 1637-1640, 2007.
2. Ishikawa, K., Nagase, T., Suyama, M., Miyajima, N., Tanaka, A., Kotani, H., Nomura, N., Ohara, O. Prediction of the coding sequences of unidentified human genes. X. The complete sequences of 100 new cDNA clones from brain which can code for large proteins in vitro. DNA Res. 5: 169-176, 1998.
3. Kolas, N. K., Chapman, J. R., Nakada, S., Ylanko, J., Chahwan, R., Sweeney, F. D., Panier, S., Mendez, M., Wildenhain, J., Thomson, T. M., Pelletier, L., Jackson, S. P., Durocher, D. Orchestration of the DNA-damage response by the RNF8 ubiquitin ligase. Science 318: 1637-1640, 2007.
NCBI and Uniprot Product Information
NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
Molecular Weight
55,517 Da
NCBI Official Full Name
E3 ubiquitin-protein ligase RNF8
NCBI Official Synonym Full Names
ring finger protein 8
NCBI Official Symbol
Rnf8
NCBI Official Synonym Symbols
AIP37; 3830404E21Rik
NCBI Protein Information
E3 ubiquitin-protein ligase RNF8; laXp180; ActA binding protein 2; actA-interacting protein 37
UniProt Protein Name
E3 ubiquitin-protein ligase RNF8
Protein Family
UniProt Gene Name
Rnf8
UniProt Synonym Gene Names
AIP37
UniProt Entry Name
RNF8_MOUSE
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Product Notes
The Rnf8 rnf8 (Catalog #AAA1753365) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-RNF8 Antibody reacts with Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's RNF8 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.1-0.25ug/ml|Mouse, Rat| FC/FACS/FCM: 1-3ug/1x106 cells|Mouse| Direct ELISA: 0.1-0.5ug/ml|Mouse|. Researchers should empirically determine the suitability of the Rnf8 rnf8 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 "RNF8, 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.Item has been added to Shopping Cart
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