XRCC4 Antibody [N17K6] | Primary Antibodies

Application: Reactivity: Mouse, Rat, Human

Usage Information

Dilution
WB1:100- 1:1000 IP1:200-1:400 IHC1:50-1:500 IF1:50-1:500

Application
WB, IP, IHC, IF, ELISA

Reactivity
Mouse, Rat, Human

Source
Mouse Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
55 kDa

Positive Control	Human breast tissue; HeLa cells; HCT-116 cells; T-47D cells; MOLT-4 cells; IB4 cells; A-431 cells
Negative Control

Datasheet & SDS

Biological Description

Specificity
XRCC4 Antibody [N17K6] detects endogenous levels of total XRCC4 protein.

Clone
N17K6

Synonym(s)
DNA repair protein XRCC4; Hxrcc4; X-ray repair cross-complementing protein 4; XRCC4

Background
XRCC4 is a core scaffolding protein in the non-homologous end joining (NHEJ) pathway, crucial for repairing DNA double-strand breaks (DSBs) caused by ionizing radiation, replication stress, or during V(D)J recombination in immune gene diversification. Unlike homologous recombination (HR), which is restricted to the S and G2 phases, XRCC4-dependent NHEJ operates throughout the entire cell cycle. XRCC4 assembles into a dumbbell-shaped tetramer. Its N-terminal globular head domains contain a β-sandwich and helix-turn-helix motif, which may mediate DNA binding, while the >120 Å C-terminal α-helical stalk forms a four-helix bundle and coiled-coil that promotes dimerization, enabling efficient multivalent engagement of DNA ends. XRCC4 tightly binds DNA ligase IV via its C-terminal stalk, displacing one XRCC4 dimer to precisely position the ligase for phosphodiester bond formation. This interaction stimulates ligase IV readenylation and catalysis, ensuring effective DSB end joining. XRCC4 also recruits XLF to form filaments that bridge and align DNA breaks, and interacts with end-processing enzymes such as PNKP through its phosphorylated C-terminal FHA-binding motifs. This facilitates the removal of 3'-phosphate and 5'-hydroxyl groups, coordinating the final ligation step of NHEJ. XRCC4 also enhances PNKP kinase/phosphatase turnover for efficient DNA end processing, and its dynamic conformational changes support complex assembly and function. Defects in XRCC4 are linked to microcephaly, growth retardation, radiosensitivity, and increased cancer risk (including hepatocellular carcinoma.

Background

XRCC4 is a core scaffolding protein in the non-homologous end joining (NHEJ) pathway, crucial for repairing DNA double-strand breaks (DSBs) caused by ionizing radiation, replication stress, or during V(D)J recombination in immune gene diversification. Unlike homologous recombination (HR), which is restricted to the S and G2 phases, XRCC4-dependent NHEJ operates throughout the entire cell cycle. XRCC4 assembles into a dumbbell-shaped tetramer. Its N-terminal globular head domains contain a β-sandwich and helix-turn-helix motif, which may mediate DNA binding, while the >120 Å C-terminal α-helical stalk forms a four-helix bundle and coiled-coil that promotes dimerization, enabling efficient multivalent engagement of DNA ends. XRCC4 tightly binds DNA ligase IV via its C-terminal stalk, displacing one XRCC4 dimer to precisely position the ligase for phosphodiester bond formation. This interaction stimulates ligase IV readenylation and catalysis, ensuring effective DSB end joining. XRCC4 also recruits XLF to form filaments that bridge and align DNA breaks, and interacts with end-processing enzymes such as PNKP through its phosphorylated C-terminal FHA-binding motifs. This facilitates the removal of 3'-phosphate and 5'-hydroxyl groups, coordinating the final ligation step of NHEJ. XRCC4 also enhances PNKP kinase/phosphatase turnover for efficient DNA end processing, and its dynamic conformational changes support complex assembly and function. Defects in XRCC4 are linked to microcephaly, growth retardation, radiosensitivity, and increased cancer risk (including hepatocellular carcinoma.

References
https://pubmed.ncbi.nlm.nih.gov/11080143/ https://pubmed.ncbi.nlm.nih.gov/28453785/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%