RAG2 Monoclonal antibody

RAG2 Monoclonal Antibody for WB, IF/ICC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG1

Reactivity

Human, pig

Applications

WB, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

1B4E5

Cat no : 66998-1-Ig

Synonyms

V(D)J recombination-activating protein 2, RAG-2, RAG 2, 1B4E5

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Validation Data Gallery

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  WB analysis using 66998-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66998-1-Ig (RAG2 antibody) at dilution of 1:4000 incubated at room temperature for 1.5 hours.

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  IF Staining of HepG2 using 66998-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using RAG2 antibody (66998-1-Ig, Clone: 1B4E5 ) at dilution of 1:800 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

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  FC experiment of HepG2 using 66998-1-Ig

  1x10^6 HepG2 cells were intracellularly stained with 0.4 ug Anti-Human RAG2 (66998-1-Ig, Clone:1B4E5) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG1 isotype control Mouse McAb (66360-1-Ig, Clone: 1F8D3) (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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Tested Applications

• Positive WB detected in: Jurkat cells, Raji cells, Ramos cells
• Positive IF/ICC detected in: HepG2 cells
• Positive FC (Intra) detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:8000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:400-1:1600
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

66998-1-Ig targets RAG2 in WB, IF/ICC, FC (Intra), ELISA applications and shows reactivity with Human, pig samples.

• Tested Reactivity: Human, pig
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: RAG2 fusion protein Ag16780
• Full Name: recombination activating gene 2
• Calculated Molecular Weight: 527 aa, 59 kDa
• Observed Molecular Weight: 57-62 kDa
• GenBank Accession Number: BC022397
• Gene Symbol: RAG2
• Gene ID (NCBI): 5897
• RRID: AB_2882315
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Recombination activating gene 2(RAG2) is core part of the RAG complex(RAG1 and RAG2), which mediates the DNA cleavage phase during V(D)J recombination. 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