RYK Rabbit Polyclonal Antibody
Catalog# HA500416
RYK Rabbit Polyclonal Antibody
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WB
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IF-Cell
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
RYK Rabbit Polyclonal Antibody
Antibody Type
Rabbit Polyclonal Antibody
Immunogen
Recombinant protein within human RYK aa 300-550.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, FC
Molecular Weight
Predicted band size: 68 kDa
Positive Control
SH-SY5Y cell lysate, NIH/3T3 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, MCF-7 cell lysates, NIH/3T3, SH-SY5Y.
Conjugation
unconjugated
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Immunogen affinity purified.
Application Dilution
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WB
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1:500-1:10,000
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IF-Cell
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1:200
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FC
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1:500-1:1,000
Target
Function
The protein encoded by this gene is an atypical member of the family of growth factor receptor protein tyrosine kinases, differing from other members at a number of conserved residues in the activation and nucleotide binding domains. This gene product belongs to a subfamily whose members do not appear to be regulated by phosphorylation in the activation segment. It has been suggested that mediation of biological activity by recruitment of a signaling-competent auxiliary protein may occur through an as yet uncharacterized mechanism. The encoded protein has a leucine-rich extracellular domain with a WIF-type Wnt binding region, a single transmembrane domain, and an intracellular tyrosine kinase domain. This protein is involved in stimulating Wnt signaling pathways such as the regulation of axon pathfinding. Alternative splicing results in multiple transcript variants encoding distinct isoforms. May be a coreceptor along with FZD8 of Wnt proteins, such as WNT1, WNT3, WNT3A and WNT5A. Involved in neuron differentiation, axon guidance, corpus callosum establishment and neurite outgrowth. In response to WNT3 stimulation, receptor C-terminal cleavage occurs in its transmembrane region and allows the C-terminal intracellular product to translocate from the cytoplasm to the nucleus where it plays a crucial role in neuronal development.
Background References
1. Lu W., Yamamoto V., Ortega B., Baltimore D. Mammalian Ryk is a Wnt coreceptor required for stimulation of neurite outgrowth. Cell 119:97-108(2004)
2. Katso R.M., Russell R.B., Ganesan T.S. Functional analysis of H-Ryk, an atypical member of the receptor tyrosine kinase family. Mol. Cell. Biol. 19:6427-6440(1999)
Subcellular Location
Cytoplasm, Membrane, Nucleus.
Synonyms
D3S3195 antibody
ERK 3 antibody
Hydroxyaryl protein kinase antibody
JTK 5 antibody
JTK 5A antibody
JTK5 antibody
JTK5A antibody
JTK5A protein tyrosine kinase antibody
receptor like tyrosine kinase antibody
receptor like tyrosine kinase precursor antibody
ExpandImages
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Western blot analysis of RYK on different lysates with Rabbit anti-RYK antibody (HA500416) at 1/500 dilution.
Lane 1: SH-SY5Y cell lysate (10 µg/Lane)
Lane 2: NIH/3T3 cell lysate (10 µg/Lane)
Lane 3: Mouse brain tissue lysate (20 µg/Lane)
Lane 4: Rat brain tissue lysate (20 µg/Lane)
Predicted band size: 68 kDa
Observed band size: 52 kDa (C terminal fragment)
Exposure time: 2 minutes;
8% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA500416) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of RYK on MCF-7 cell lysates with Rabbit anti-RYK antibody (HA500416) at 1/10,000 dilution.
Lysates/proteins at 10 µg/Lane.
Predicted band size: 68 kDa
Observed band size: 42 kDa (intracellular domain)
Exposure time: 1 minute;
10% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA500416) at 1/10,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of NIH/3T3 cells labeling RYK with Rabbit anti-RYK antibody (HA500416) at 1/200 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-RYK antibody (HA500416) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. -
Flow cytometric analysis of NIH/3T3 cells labeling RYK.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA500416, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Flow cytometric analysis of SH-SY5Y cells labeling RYK.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA500416, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"