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MABE1104

Sigma-Aldrich

Anti-RAG-2 Antibody, clone 39

culture supernatant, from rabbit

Synonym(s):

V(D)J recombination-activating protein 2, RAG-2

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

rabbit

Quality Level

antibody form

culture supernatant

antibody product type

primary antibodies

clone

monoclonal

species reactivity

mouse

packaging

antibody small pack of 25 μL

technique(s)

ChIP: suitable
immunofluorescence: suitable
immunoprecipitation (IP): suitable
western blot: suitable

UniProt accession no.

target post-translational modification

unmodified

General description

V(D)J recombination-activating protein 2 (UniProt: P21784; also known as RAG-2) is encoded by the Rag2 (also known as Rag-2) gene (Gene ID: 19374) in murine species. RAG-2 is a nuclear protein that is a core component of the multiprotein RAG complex. It is predominantly found in maturing lymphoid cells. RAG complex mediates the DNA cleavage phase during V(D)J recombination. During this phase 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. DNA cleavage by the RAG complex is reported to occur by first introducing a nick 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 trans-esterification reaction, thereby creating 4 DNA ends: 2 hairpin coding ends and 2 blunt, 5′-phosphorylated ends. The chromatin structure is shown to play a critical role in the V(D)J recombination reactions and the presence of histone H3 trimethylated at Lys4 (H3K4me3) stimulates both the nicking and hairpinning steps. RAG-2 contains an atypical PHD-type zinc finger (a 416-484) that recognizes and binds histone H3 trimethylated on Lys4 (H3K4me3). 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. Although RAG-2 is not the catalytic component, it is required for all known catalytic activities mediated by RAG1. Mice with defective Rag2 gene fail to produce mature B or T-lymphocytes.

Specificity

Clone 39 is a rabbit monoclonal antibody that detects RAG-2 protein in murine species.

Immunogen

GST-tagged recombinantn fragment corresponind to the first 490 amino acids of murine V(D)J recombination-activating protein 2 (RAG-2).

Application

Detect V(D)J recombination-activating protein 2 using this rabbit monoclonal Anti-RAG-2, clone 39, Cat. No. MABE1104-25UL, validated for use in Western Blotting and Chromatin Immunoprecipitation (ChIP), Immunofluorescence and Immunoprecipitation.
Immunofluorescence Analysis: A representative lot detected RAG-2 in Immunofluorescence applications (Chan, E.A., et. al. (2013). Proc Natl Acad Sci USA. 110(48):E4628-37; Leu, T.M., et. al. (1995). Mol Cell Biol. 15(10):5657-70).

Immunoprecipitation Analysis: A representative lot detected RAG-2 in Immunoprecipitation applications (Leu, T.M., et. al. (1995). Mol Cell Biol. 15(10):5657-70).

Chromatin Immunoprecipitation (ChIP) Analysis: A representative lot detected RAG-2 in Chromatin Immunoprecipitation applications (Dose, M., et. al. (2014). Proc Natl Acad Sci USA. 111(1):391-6; Ji, Y., et. al. (2010). Cell. 141(3):419-31).

Western Blotting Analysis: A representative lot detected RAG-2 in Western Blotting applications (Coster, G., et. al. (2012). J Biol Chem. 287(43):36488-98; Leu, T.M., et. al. (1995). Mol Cell Biol. 15(10):5657-70; Dose, M., et. al. (2014). Proc Natl Acad Sci USA. 111(1):391-6).
Research Category
Apoptosis & Cancer

Quality

Evaluated by Western Blotting in mouse bone marrow tissue lysate.
Western Blotting Analysis: A 1:2,000 dilution of this antibody detected RAG-2 in 10 µg of mouse bone marrow tissue lysate.

Physical form

Rabbit monoclonal antibody in tissue culture supernatant without preservatives.
Unpurified

Storage and Stability

Stable for 1 year at -20°C from date of receipt.

Other Notes

Concentration: Please refer to lot specific datasheet.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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Storage Class Code

10 - Combustible liquids

WGK

WGK 1


Certificates of Analysis (COA)

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N Max Schabla et al.
Journal of immunology (Baltimore, Md. : 1950), 201(3), 930-939 (2018-06-22)
The assembly of Ig genes in developing B lymphocytes by V(D)J recombination is initiated by the RAG1-RAG2 endonuclease complex. We previously identified an interaction between RAG1 and viral protein R binding protein (VprBP) (also known as DNA damage binding protein

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