Anti-RAG2 Antibody (CAB5626)

Antibody Name:	Anti-RAG2 Antibody [KO Validated]
Antibody SKU:	CAB5626
Antibody Size:	20uL, 50uL, 100uL
Application:	WB IF
Reactivity:	Human, Mouse
Host Species:	Rabbit
Immunogen:	Recombinant fusion protein containing a sequence corresponding to amino acids 258-527 of human RAG2 (NP_001230715.1).

Application:	WB IF
Recommended Dilution:	WB 1:500 - 1:2000 IF 1:50 - 1:200
Reactivity:	Human, Mouse
Positive Samples:	Raji, Jurkat, 293T, HepG2

Immunogen:	Recombinant fusion protein containing a sequence corresponding to amino acids 258-527 of human RAG2 (NP_001230715.1).
Purification Method:	Affinity purification
Storage Buffer:	Store at -20'C. Avoid freeze / thaw cycles. Buffer: PBS with 0.02% sodium azide, 50% glycerol, pH7.3.
Isotype:	IgG
Sequence:	VSSA ILTQ TNND EFVI VGGY QLEN QKRM ICNI ISLE DNKI EIRE METP DWTP DIKH SKIW FGSN MGNG TVFL GIPG DNKQ VVSE GFYF YMLK CAED DTNE EQTT FTNS QTST EDPG DSTP FEDS EEFC FSAE ANSF DGDD EFDT YNED DEED ESET GYWI TCCP TCDV DINT WVPF YSTE LNKP AMIY CSHG DGHW VHAQ CMDL AERT LIHL SAGS NKYY CNEH VEIA RALH TPQR VLPL KKPP MKSL RKKG SGKI LTPA KKSF LRRL FD
Gene ID:	5897
Uniprot:	P55895
Cellular Location:	Nucleus
Calculated MW:	59kDa
Observed MW:	59kDa

Synonyms:

RAG2, RAG-2

Background:

This gene encodes a protein that is involved in the initiation of V(D)J recombination during B and T cell development. This protein forms a complex with the product of the adjacent recombination activating gene 1, and this complex can form double-strand breaks by cleaving DNA at conserved recombination signal sequences. The recombination activating gene 1 component is thought to contain most of the catalytic activity, while the N-terminal of the recombination activating gene 2 component is thought to form a six-bladed propeller in the active core that serves as a binding scaffold for the tight association of the complex with DNA. A C-terminal plant homeodomain finger-like motif in this protein is necessary for interactions with chromatin components, specifically with histone H3 that is trimethylated at lysine 4. Mutations in this gene cause Omenn syndrome, a form of severe combined immunodeficiency associated with autoimmune-like symptoms.

UniProt Protein Function:	RAG2: Core 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. DNA cleavage by the RAG complex 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 the RAG complex, RAG2 is not the catalytic component but is required for all known catalytic activities mediated by RAG1. It probably acts as a sensor of chromatin state that recruits the RAG complex to H3K4me3. Component of the RAG complex composed of core components RAG1 and RAG2, and associated component HMGB1 or HMGB2. Cells of the B- and T-lymphocyte lineages. Belongs to the RAG2 family.
UniProt Protein Details:	Protein type:Enzyme, misc. Chromosomal Location of Human Ortholog: 11p13 Cellular Component: nucleoplasm Molecular Function:chromatin binding; methylated histone residue binding; phosphatidylinositol-3,4,5-triphosphate binding; phosphatidylinositol-3,4-bisphosphate binding; phosphatidylinositol-4,5-bisphosphate binding; phosphoinositide binding; zinc ion binding Biological Process: B cell differentiation; pre-B cell allelic exclusion; T cell differentiation in the thymus; V(D)J recombination Disease: Combined Cellular And Humoral Immune Defects With Granulomas; Omenn Syndrome; Severe Combined Immunodeficiency, Autosomal Recessive, T Cell-negative, B Cell-negative, Nk Cell-positive
NCBI Summary:	This gene encodes a protein that is involved in the initiation of V(D)J recombination during B and T cell development. This protein forms a complex with the product of the adjacent recombination activating gene 1, and this complex can form double-strand breaks by cleaving DNA at conserved recombination signal sequences. The recombination activating gene 1 component is thought to contain most of the catalytic activity, while the N-terminal of the recombination activating gene 2 component is thought to form a six-bladed propeller in the active core that serves as a binding scaffold for the tight association of the complex with DNA. A C-terminal plant homeodomain finger-like motif in this protein is necessary for interactions with chromatin components, specifically with histone H3 that is trimethylated at lysine 4. Mutations in this gene cause Omenn syndrome, a form of severe combined immunodeficiency associated with autoimmune-like symptoms. [provided by RefSeq, Jul 2008]
UniProt Code:	P55895
NCBI GenInfo Identifier:	2498830
NCBI Gene ID:	5897
NCBI Accession:	P55895.1
UniProt Secondary Accession:	P55895,Q8TBL4, A8K9E9,
UniProt Related Accession:	P55895
Molecular Weight:	59,241 Da
NCBI Full Name:	V(D)J recombination-activating protein 2
NCBI Synonym Full Names:	recombination activating gene 2
NCBI Official Symbol:	RAG2
NCBI Official Synonym Symbols:	RAG-2
NCBI Protein Information:	V(D)J recombination-activating protein 2
UniProt Protein Name:	V(D)J recombination-activating protein 2
Protein Family:	Seed allergenic protein
UniProt Gene Name:	RAG2
UniProt Entry Name:	RAG2_HUMAN

	Western blot analysis of extracts of various cell lines, using RAG2 antibody (CAB5626) at 1:1000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (CABS014) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit (RM00020). Exposure time: 90s.
	Western blot analysis of extracts from normal (control) and RAG2 Rabbit pAb knockout (KO) 293T cells, using RAG2 Rabbit pAb antibody (CAB5626) at 1:500 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (CABS014) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit (RM00020). Exposure time: 1s.
	Immunofluorescence analysis of U2OS cells using RAG2 antibody (CAB5626). Blue: DAPI for nuclear staining.
	Immunofluorescence analysis of GFP-RNF168 transgenic U2OS cells using RAG2 antibody (CAB5626). (Green)GFP-RNF168 fusion protein expression for DNA damage marker.Blue: DAPI for nuclear staining.RNF168(GFP) can be used to mark cells damaged by UV-A laser for they always gather around DNA damage region.

Antibodies	ELISA Kits
Anti-RAG2 Antibody (CAB12488)	Human RAG2 ELISA Kit
	Human V(D)J recombination-activating protein 2 (RAG2) ELISA Kit

Secondary Antibody

Anti-HRP Goat Anti-Rabbit IgG (H+L) Antibody (CABS014)