RAG2 Polyclonal Antibody

货号：PA5-22048

规格：100 µL

价格：5547

产品类型：标签抗体

品牌：Thermo Fisher

抗原：Recombinant fragment corresponding to a region wit

物种：其它

宿主：兔

抗体亚型：IgG

荧光染料：其它

类型:	一抗	同型对照：
浓度：	0.37mg/mL	用法：	1:100-1:1000（ICC）；1:100-1:1000(IF);1:100-1:1000(IHC(P)）;1:500-1:3000(WB)

产品详细信息Recommended positive controls: Molt-4, rat thymus.Predicted reactivity: Mouse (85%), Rat (86%), Pig (90%), Rabbit (91%), Bovine (91%).Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.靶标信息RAG2 (V(D)J recombination-activating protein 2) is a 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: 1) first a 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 are created. The chromatin structure plays an essential rold in the V(D)J recombination reactions and the presence of histone H3 trimethylated at 'Lys-4' (H3K4me3) stimulates both the nicking and hair pinning 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. This prevents 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. Mutations affecting the gene can results in combined cellular and humoral immune defects with granulomas, severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-negative/NK-cell-positive, and Omenn syndrome.

数据：

RAG2 Antibody (PA5-22048) in IFImmunofluorescence analysis of RAG2 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with RAG2 Polyclonal Antibody (Product # PA5-22048) at 1:200 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominant nuclear localization along with weak cytoplasmic signal. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification. RAG2 Antibody (PA5-22048) in IHC (P)Immunohistochemical analysis of RAG2 in paraffin-embedded DLD-1 xenograft using a RAG2 polyclonal antibody (Product # PA5-22048) at a dilution of 1:100. RAG2 Antibody (PA5-22048) in WBWestern blot analysis was performed on modified whole cell extracts (1% SDS) (30 µg lysate) of Jurkat (Lane 1), Ramos (Lane 2), Hep G2 (Lane 3), HeLa (Lane 4) and MOLT-4 (Lane 5). The blot was probed with Anti-RAG2 Polyclonal Antibody (Product # PA5-22048, 1:500 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 55 kDa band corresponding to RAG2 was observed in all cell lines along with additional uncharacterized bands (*) between ~70 kDa and ~110 kDa.

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