CUL3 Antibody (Rabbit mAb) [N12N7]

Application: Reactivity: Human, Mouse, Rat, Monkey

Usage Information

Dilution
WB1:1000-1:5000 IP1:50

Application
WB, IP

Reactivity
Human, Mouse, Rat, Monkey

Source
Rabbit 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 Observed MW
89 kDa N/A
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight. Post-translational modifications(e.g., phosphorylation, glycosylation); Splice variants and isoforms; Relative charge; Multimerization.

Positive Control	A-204 cells; A-673 cells; Neuro-2a cells; C6 cells; COS-7 cells; K-562 cells (MLN4924, 0.3uM, 16 h)
Negative Control

Datasheet & SDS

Biological Description

Specificity
CUL3 Antibody (Rabbit mAb) [N12N7] detects endogenous levels of total CUL3 protein.

Clone
N12N7

Synonym(s)
CUL-3; CUL3; cullin 3; Cullin-3; KIAA0617; PHA2E

Background
CUL3 (Cullin-3) belongs to the cullin family of scaffold proteins that assemble Cullin-RING ubiquitin ligase (CRL) complexes, the largest class of E3 ligases in eukaryotic cells, and CUL3 forms one of eight distinct cullin-based assemblies distinguished by their substrate-recognition modules. CUL3 anchors the RING-finger catalytic protein Rbx1 at its C-terminus, positioning the ubiquitin-charged E2 conjugating enzyme for substrate transfer, while its N-terminal domain binds directly to BTB (Bric-a-brac/Tramtrack/Broad complex) domain-containing adaptor proteins that simultaneously serve as substrate receptors, a modular arrangement that removes the need for a separate linker subunit and distinguishes CUL3 complexes from other cullin-RING assemblies. BTB adaptors combine their conserved Cul3-binding domain with a second, variable domain such as Kelch, MATH, zinc finger, or PHR that dictates substrate specificity, and this combinatorial pairing allows CUL3 to organize a large repertoire of distinct ubiquitin ligase complexes from a single scaffold. BTB adaptors are also capable of dimerization, allowing two CUL3 molecules to be incorporated into a single ligase complex, a configuration linked to substrate ubiquitination efficiency. Ligase activation depends on conjugation of the ubiquitin-like protein Nedd8 onto a conserved C-terminal lysine of CUL3, which reshapes the Rbx1 RING domain and increases its conformational flexibility, bringing the E2 enzyme into proximity with the substrate for efficient ubiquitin transfer. Through this mechanism, CUL3 complexes direct proteasomal degradation of substrates governing cell cycle progression, transcriptional regulation, and developmental differentiation. Among characterized CUL3-BTB pairings, Keap1 links CUL3 to Nrf2, controlling the oxidative stress response, KLHL20 bridges CUL3 to substrates including PML and DAPK, influencing tumor suppressor turnover, and SPOP directs CUL3 activity toward substrates involved in transcriptional and signaling regulation, and dysregulation of these adaptor-substrate relationships is observed across multiple human cancers, altering ligase output and stabilizing or depleting proteins that shape tumor progression and therapeutic response. The single-adaptor bridging design, combined with tissue-specific expression of individual BTB proteins, makes CUL3 ligase complexes a tractable framework for dissecting selective protein degradation pathways and for identifying which adaptor-substrate module underlies a particular physiological or disease phenotype, information that is directly relevant when researchers are selecting CUL3-associated targets for functional or therapeutic study.

Background

CUL3 (Cullin-3) belongs to the cullin family of scaffold proteins that assemble Cullin-RING ubiquitin ligase (CRL) complexes, the largest class of E3 ligases in eukaryotic cells, and CUL3 forms one of eight distinct cullin-based assemblies distinguished by their substrate-recognition modules. CUL3 anchors the RING-finger catalytic protein Rbx1 at its C-terminus, positioning the ubiquitin-charged E2 conjugating enzyme for substrate transfer, while its N-terminal domain binds directly to BTB (Bric-a-brac/Tramtrack/Broad complex) domain-containing adaptor proteins that simultaneously serve as substrate receptors, a modular arrangement that removes the need for a separate linker subunit and distinguishes CUL3 complexes from other cullin-RING assemblies. BTB adaptors combine their conserved Cul3-binding domain with a second, variable domain such as Kelch, MATH, zinc finger, or PHR that dictates substrate specificity, and this combinatorial pairing allows CUL3 to organize a large repertoire of distinct ubiquitin ligase complexes from a single scaffold. BTB adaptors are also capable of dimerization, allowing two CUL3 molecules to be incorporated into a single ligase complex, a configuration linked to substrate ubiquitination efficiency. Ligase activation depends on conjugation of the ubiquitin-like protein Nedd8 onto a conserved C-terminal lysine of CUL3, which reshapes the Rbx1 RING domain and increases its conformational flexibility, bringing the E2 enzyme into proximity with the substrate for efficient ubiquitin transfer. Through this mechanism, CUL3 complexes direct proteasomal degradation of substrates governing cell cycle progression, transcriptional regulation, and developmental differentiation. Among characterized CUL3-BTB pairings, Keap1 links CUL3 to Nrf2, controlling the oxidative stress response, KLHL20 bridges CUL3 to substrates including PML and DAPK, influencing tumor suppressor turnover, and SPOP directs CUL3 activity toward substrates involved in transcriptional and signaling regulation, and dysregulation of these adaptor-substrate relationships is observed across multiple human cancers, altering ligase output and stabilizing or depleting proteins that shape tumor progression and therapeutic response. The single-adaptor bridging design, combined with tissue-specific expression of individual BTB proteins, makes CUL3 ligase complexes a tractable framework for dissecting selective protein degradation pathways and for identifying which adaptor-substrate module underlies a particular physiological or disease phenotype, information that is directly relevant when researchers are selecting CUL3-associated targets for functional or therapeutic study.

References
https://pubmed.ncbi.nlm.nih.gov/31898230/ https://pubmed.ncbi.nlm.nih.gov/27200299/

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%