CRX Antibody (Rabbit mAb) [F1B1]

Application: Reactivity: Human, Mouse, Rat

Usage Information

Dilution
WB1:1000 IHC1:250 - 1:500

Application
WB, IHC

Reactivity
Human, Mouse, Rat

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
32 kDa 37 kDa
^*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.

Datasheet & SDS

Biological Description

Specificity
CRX Antibody (Rabbit mAb) [F1B1] detects endogenous levels of total CRX protein.

Clone
F1B1

Synonym(s)
CORD2, CRX, Cone-rod homeobox protein

Background
Cone-rod homeobox (CRX) is a retina-specific K50 homeodomain transcription factor of the Otx family that binds ATTA-containing cis-regulatory motifs such as 5′‑TAATCC/A‑3′ upstream of photoreceptor genes and drives the differentiation, maturation and maintenance of rod and cone photoreceptors in the mammalian retina. The protein contains a highly conserved homeodomain that mediates sequence-specific DNA binding and additional transactivation domains that recruit co-regulators to photoreceptor promoters and enhancers, enabling CRX to act as a central node in the transcription factor network that controls photoreceptor identity. CRX is one of the earliest photoreceptor-enriched transcription factors expressed during retinal development and binds thousands of sites across the genome; chromatin immunoprecipitation combined with functional assays show that a subset of these sites correspond to regulatory elements at key phototransduction genes, including opsins, transducin, arrestin and components of the ribbon synapse, where CRX occupancy is required for high-level, cell type–specific transcription. At many photoreceptor gene promoters, CRX acts synergistically with rod and cone determinant factors such as NRL, NR2E3, RORβ and RXRγ, forming cell type–specific complexes in which pioneer factors and nuclear receptors cooperate with CRX to fine-tune expression of rod- versus cone-specific programs and to maintain subtype identity in the mature retina. Genome-wide analysis of CRX binding and transcriptional output reveals that CRX accelerates chromatin opening at photoreceptor regulatory elements during development, increasing accessibility at enhancers and promoters that subsequently acquire active histone marks, demonstrating a role for CRX in coordinating epigenetic remodeling with transcriptional activation in differentiating photoreceptors. Structural and biochemical studies of the human CRX homeodomain show that disease-associated missense mutations alter DNA binding affinity, specificity or transactivation capacity, and patient-derived mutations cluster in regions critical for homeodomain structure and cofactor interactions, providing mechanistic explanations for the retinal dystrophies linked to CRX variation. In mouse models with Crx loss-of-function, photoreceptor cells fail to properly express opsins and other phototransduction components, exhibit defective outer segment formation and ribbon synapse assembly, and undergo progressive degeneration, confirming that CRX-dependent transcription is essential both for initial photoreceptor differentiation and for long-term survival and function. Together, these findings describe CRX as an Otx-like homeobox transcription factor whose structured homeodomain recognizes photoreceptor cis-elements, whose partnerships with NRL, RORβ, NR2E3 and related regulators define rod and cone transcriptional circuits, and whose activity drives chromatin accessibility, phototransduction gene expression and presynaptic active zone organization, establishing CRX as a pivotal target for studies of retinal development and inherited photoreceptor degeneration.

Background

Cone-rod homeobox (CRX) is a retina-specific K50 homeodomain transcription factor of the Otx family that binds ATTA-containing cis-regulatory motifs such as 5′‑TAATCC/A‑3′ upstream of photoreceptor genes and drives the differentiation, maturation and maintenance of rod and cone photoreceptors in the mammalian retina. The protein contains a highly conserved homeodomain that mediates sequence-specific DNA binding and additional transactivation domains that recruit co-regulators to photoreceptor promoters and enhancers, enabling CRX to act as a central node in the transcription factor network that controls photoreceptor identity. CRX is one of the earliest photoreceptor-enriched transcription factors expressed during retinal development and binds thousands of sites across the genome; chromatin immunoprecipitation combined with functional assays show that a subset of these sites correspond to regulatory elements at key phototransduction genes, including opsins, transducin, arrestin and components of the ribbon synapse, where CRX occupancy is required for high-level, cell type–specific transcription. At many photoreceptor gene promoters, CRX acts synergistically with rod and cone determinant factors such as NRL, NR2E3, RORβ and RXRγ, forming cell type–specific complexes in which pioneer factors and nuclear receptors cooperate with CRX to fine-tune expression of rod- versus cone-specific programs and to maintain subtype identity in the mature retina. Genome-wide analysis of CRX binding and transcriptional output reveals that CRX accelerates chromatin opening at photoreceptor regulatory elements during development, increasing accessibility at enhancers and promoters that subsequently acquire active histone marks, demonstrating a role for CRX in coordinating epigenetic remodeling with transcriptional activation in differentiating photoreceptors. Structural and biochemical studies of the human CRX homeodomain show that disease-associated missense mutations alter DNA binding affinity, specificity or transactivation capacity, and patient-derived mutations cluster in regions critical for homeodomain structure and cofactor interactions, providing mechanistic explanations for the retinal dystrophies linked to CRX variation. In mouse models with Crx loss-of-function, photoreceptor cells fail to properly express opsins and other phototransduction components, exhibit defective outer segment formation and ribbon synapse assembly, and undergo progressive degeneration, confirming that CRX-dependent transcription is essential both for initial photoreceptor differentiation and for long-term survival and function. Together, these findings describe CRX as an Otx-like homeobox transcription factor whose structured homeodomain recognizes photoreceptor cis-elements, whose partnerships with NRL, RORβ, NR2E3 and related regulators define rod and cone transcriptional circuits, and whose activity drives chromatin accessibility, phototransduction gene expression and presynaptic active zone organization, establishing CRX as a pivotal target for studies of retinal development and inherited photoreceptor degeneration.

References
https://pubmed.ncbi.nlm.nih.gov/17662965/ https://pubmed.ncbi.nlm.nih.gov/35255174/

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%