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RNF74 Polyclonal Antibody, RBITC Conjugated

Applications

  • WB
  • FCM
  • IF(IHC-P)
  • IF(IHC-F)
  • IF(ICC)

Reactivity

  • Mouse

Predicted Reactivity

  • Human
  • Rat
  • Sheep
  • Pig
Overview
Catalog # bs-6941R-RBITC-100uL
Product Name RNF74 Polyclonal Antibody, RBITC Conjugated
Applications
WB FCM IF(IHC-P) IF(IHC-F) IF(ICC)
Reactivity Mouse
Predicted Reactivity Human, Rat, Sheep, Pig
Specifications
Conjugation RBITC
Host Rabbit
Source KLH conjugated synthetic peptide derived from human RAG1/RNF74
Clonality Polyclonal
Isotype IgG
Concentration 1ug/ul
Purification Purified by Protein A.
Storage Buffer Aqueous buffered solution containing 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition Store at -20C. Aliquot into multiple vials to avoid repeated freeze-thaw cycles.
Target
Gene ID 5896
Swiss Prot P15918
Subcellular location Nucleus
Synonyms RAG-1; RNF74; V(D)J recombination-activating protein 1; RING finger protein 74; RAG1
Background 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.
Application Dilution
WB 1:300-5000
FCM 1:20-100
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200

Frequently asked questions

Which applications is RNF74 Polyclonal Antibody, RBITC Conjugated (bs-6941R-RBITC) validated for, and at what dilution?

bs-6941R-RBITC is validated for WB (1:300-5000); FCM (1:20-100); IF(IHC-P) (1:50-200); IF(IHC-F) (1:50-200); IF(ICC) (1:50-200). These are recommended starting dilutions; optimal dilutions should be determined by the end user.

Which species does RNF74 Polyclonal Antibody, RBITC Conjugated (bs-6941R-RBITC) react with?

Tested reactivity: Mouse. Predicted reactivity (expected from sequence homology but not yet tested): Human, Rat, Sheep, Pig.

How should bs-6941R-RBITC be stored?

Store at -20C. Aliquot into multiple vials to avoid repeated freeze-thaw cycles. Storage buffer: Aqueous buffered solution containing 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.

What immunogen was used to raise RNF74 Polyclonal Antibody, RBITC Conjugated (bs-6941R-RBITC)?

KLH conjugated synthetic peptide derived from human RAG1/RNF74

What are the excitation and emission wavelengths of the RBITC conjugate (bs-6941R-RBITC)?

RBITC on bs-6941R-RBITC: excitation / emission 540nm/625nm.

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