Western Blot (WB)
(Figure 1. Western blot analysis of RAG1 using anti-RAG1 antibody (MBS1753308).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: human Jurkat whole cell lysatesLane 2: human Jurkat whole cell lysatesLane 3: mouse thymus tissue 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-RAG1 antigen affinity purified polyclonal antibody (Catalog # MBS1753308) at 0. 5 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 1:5000 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 RAG1 at approximately 120KD. The expected band size for RAG1 is at 120KD. )
Rabbit anti-Human, Mouse RAG1 Polyclonal Antibody | anti-RAG1 antibody
Anti-RAG1 Antibody
Western Blot (WB)
(Figure 1. Western blot analysis of RAG1 using anti-RAG1 antibody (MBS1753308).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: human Jurkat whole cell lysatesLane 2: human Jurkat whole cell lysatesLane 3: mouse thymus tissue 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-RAG1 antigen affinity purified polyclonal antibody (Catalog # MBS1753308) at 0. 5 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 1:5000 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 RAG1 at approximately 120KD. The expected band size for RAG1 is at 120KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of RAG1 using anti-RAG1 antibody (MBS1753308).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: human Jurkat whole cell lysatesLane 2: human Jurkat whole cell lysatesLane 3: mouse thymus tissue 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-RAG1 antigen affinity purified polyclonal antibody (Catalog # MBS1753308) at 0. 5 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 1:5000 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 RAG1 at approximately 120KD. The expected band size for RAG1 is at 120KD. )
2. Blanquet, V., Turleau, C., Goossens, M., Besmond, C. Assignment of the human recombination activating gene 1 (RAG1) to the 14q21. 3-q22. 2 region. Genomics 13: 488-489, 1992.
3. Corneo, B., Moshous, D., Gungor, T., Wulffraat, N., Philippet, P., Le Deist, F., Fischer, A., de Villartay, J. -P. Identical mutations in RAG1 or RAG2 genes leading to defective V(D)J recombinase activity can cause either T-B-severe combined immune deficiency or Omenn syndrome. Blood 97: 2772-2776, 2001.
NCBI and Uniprot Product Information
NCBI Description
The protein encoded by this gene is involved in activation of immunoglobulin V-D-J recombination. The encoded protein is involved in recognition of the DNA substrate, but stable binding and cleavage activity also requires RAG2. Defects in this gene can be the cause of several diseases. [provided by RefSeq, Jul 2008]
Uniprot Description
RAG1: Catalytic component of the RAG complex, a multiprotein complex that mediates the DNA cleavage phase during V(D)J recombination. V(D)J recombination assembles a diverse repertoire of immunoglobulin and T-cell receptor genes in developing B and T- lymphocytes through rearrangement of different V (variable), in some cases D (diversity), and J (joining) gene segments. In the RAG complex, RAG1 mediates the DNA-binding to the conserved recombination signal sequences (RSS) and catalyzes the DNA cleavage activities by introducing a double-strand break between the RSS and the adjacent coding segment. RAG2 is not a catalytic component but is required for all known catalytic activities. DNA cleavage occurs in 2 steps: a first nick is introduced in the top strand immediately upstream of the heptamer, generating a 3'- hydroxyl group that can attack the phosphodiester bond on the opposite strand in a direct transesterification reaction, thereby creating 4 DNA ends: 2 hairpin coding ends and 2 blunt, 5'- phosphorylated ends. The chromatin structure plays an essential role in the V(D)J recombination reactions and the presence of histone H3 trimethylated at 'Lys-4' (H3K4me3) stimulates both the nicking and haipinning steps. The RAG complex also plays a role in pre-B cell allelic exclusion, a process leading to expression of a single immunoglobulin heavy chain allele to enforce clonality and monospecific recognition by the B-cell antigen receptor (BCR) expressed on individual B-lymphocytes. The introduction of DNA breaks by the RAG complex on one immunoglobulin allele induces ATM-dependent repositioning of the other allele to pericentromeric heterochromatin, preventing accessibility to the RAG complex and recombination of the second allele. In addition to its endonuclease activity, RAG1 also acts as a E3 ubiquitin-protein ligase that mediates monoubiquitination of histone H3. Histone H3 monoubiquitination is required for the joining step of V(D)J recombination. Mediates polyubiquitination of KPNA1. Homodimer. Component of the RAG complex composed of core components RAG1 and RAG2, and associated component HMGB1 or HMGB2. Maturing lymphoid cells. Belongs to the RAG1 family.
Protein type: EC 3.1.-.-; EC 6.3.2.-; DNA-binding; Ligase; Ubiquitin ligase; Ubiquitin conjugating system; EC 6.3.2.19
Chromosomal Location of Human Ortholog: 11p13
Cellular Component: nucleoplasm; nucleus
Molecular Function: acid-amino acid ligase activity; DNA binding; endonuclease activity; histone binding; metal ion binding; protein binding; protein homodimerization activity; sequence-specific DNA binding; ubiquitin-protein ligase activity; zinc ion binding
Biological Process: adaptive immune response; B cell differentiation; DNA recombination; histone monoubiquitination; immune response; negative regulation of caspase activity; positive regulation of T cell differentiation; pre-B cell allelic exclusion; protein autoubiquitination; T cell differentiation in the thymus; T cell homeostasis; thymus development; V(D)J recombination
Disease: Alpha/beta T-cell Lymphopenia With Gamma/delta T-cell Expansion, Severe Cytomegalovirus Infection, And Autoimmunity; Combined Cellular And Humoral Immune Defects With Granulomas; Omenn Syndrome; Severe Combined Immunodeficiency, Autosomal Recessive, T Cell-negative, B Cell-negative, Nk Cell-positive
Research Articles on RAG1
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Product Notes
The RAG1 rag1 (Catalog #AAA1753308) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-RAG1 Antibody reacts with Human, Mouse and may cross-react with other species as described in the data sheet. AAA Biotech's RAG1 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: 0.25-0.5ug/ml|Human, Mouse|. Researchers should empirically determine the suitability of the RAG1 rag1 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 "RAG1, 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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