Product: Phospho-CXCR4 (Ser324/Ser325) Antibody
Catalog: AF3641
Description: Rabbit polyclonal antibody to Phospho-CXCR4 (Ser324/Ser325)
Application: ELISA(peptide)
Reactivity: Human, Mouse, Rat
Mol.Wt.: 40kD(Calculated).
Uniprot: P61073
RRID: AB_2846955

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Product Info

Source:
Rabbit
Application:
ELISA(peptide) 1:20000-1:40000
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Clonality:
Polyclonal
Specificity:
Phospho-CXCR4 (Ser324/Ser325) Antibody detects endogenous levels of CXCR4 only when phosphorylated at Ser324/Ser325.
RRID:
AB_2846955
Cite Format: Affinity Biosciences Cat# AF3641, RRID:AB_2846955.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

C-X-C chemokine receptor type 4; CD184; CD184 antigen; Chemokine (C X C motif) receptor 4; Chemokine CXC Motif Receptor 4; CXC-R4; CXCR-4; CXCR4; CXCR4_HUMAN; D2S201E; FB22; Fusin; HM89; HSY3RR; LAP 3; LAP3; LCR1; LESTR; Leukocyte derived seven transmembrane domain receptor; Leukocyte-derived seven transmembrane domain receptor; Lipopolysaccharide associated protein 3; Neuropeptide Y receptor Y3; NPY3R; NPYR; NPYRL; NPYY3; NPYY3R; Probable G protein coupled receptor lcr1 homolog; SDF 1 receptor; SDF-1 receptor; SEVEN-TRANSMEMBRANE-SEGMENT RECEPTOR; Stromal cell derived factor 1 receptor; Stromal cell-derived factor 1 receptor; WHIM; WHIMS;

Immunogens

Immunogen:

A synthesized peptide derived from human CXCR4 around the phosphorylation site of Ser324/Ser325.

Uniprot:
Gene(ID):
Expression:
P61073 CXCR4_HUMAN:

Expressed in numerous tissues, such as peripheral blood leukocytes, spleen, thymus, spinal cord, heart, placenta, lung, liver, skeletal muscle, kidney, pancreas, cerebellum, cerebral cortex and medulla (in microglia as well as in astrocytes), brain microvascular, coronary artery and umbilical cord endothelial cells. Isoform 1 is predominant in all tissues tested.

Sequence:
MEGISIYTSDNYTEEMGSGDYDSMKEPCFREENANFNKIFLPTIYSIIFLTGIVGNGLVILVMGYQKKLRSMTDKYRLHLSVADLLFVITLPFWAVDAVANWYFGNFLCKAVHVIYTVNLYSSVLILAFISLDRYLAIVHATNSQRPRKLLAEKVVYVGVWIPALLLTIPDFIFANVSEADDRYICDRFYPNDLWVVVFQFQHIMVGLILPGIVILSCYCIIISKLSHSKGHQKRKALKTTVILILAFFACWLPYYIGISIDSFILLEIIKQGCEFENTVHKWISITEALAFFHCCLNPILYAFLGAKFKTSAQHALTSVSRGSSLKILSKGKRGGHSSVSTESESSSFHSS

PTMs - P61073 As Substrate

Site PTM Type Enzyme
S5 Phosphorylation
N11 N-Glycosylation
Y12 Phosphorylation
S18 O-Glycosylation
S18 Phosphorylation
Y157 Phosphorylation
S178 Phosphorylation
K310 Ubiquitination
S312 Phosphorylation
T318 Phosphorylation
S319 Phosphorylation
S321 Phosphorylation
S324 Phosphorylation Q05655 (PRKCD) , P43250 (GRK6)
S325 Phosphorylation P43250 (GRK6) , Q05655 (PRKCD)
K327 Ubiquitination
S330 Phosphorylation P43250 (GRK6)
K331 Ubiquitination
K333 Ubiquitination
R334 Methylation
S338 Phosphorylation
S339 Phosphorylation P34947 (GRK5) , P11309 (PIM1) , P43250 (GRK6)
S341 Phosphorylation
T342 Phosphorylation
S344 Phosphorylation
S346 Phosphorylation
S347 Phosphorylation
S348 Phosphorylation
S351 Phosphorylation
S352 Phosphorylation

Research Backgrounds

Function:

Receptor for the C-X-C chemokine CXCL12/SDF-1 that transduces a signal by increasing intracellular calcium ion levels and enhancing MAPK1/MAPK3 activation. Involved in the AKT signaling cascade. Plays a role in regulation of cell migration, e.g. during wound healing. Acts as a receptor for extracellular ubiquitin; leading to enhanced intracellular calcium ions and reduced cellular cAMP levels. Binds bacterial lipopolysaccharide (LPS) et mediates LPS-induced inflammatory response, including TNF secretion by monocytes. Involved in hematopoiesis and in cardiac ventricular septum formation. Also plays an essential role in vascularization of the gastrointestinal tract, probably by regulating vascular branching and/or remodeling processes in endothelial cells. Involved in cerebellar development. In the CNS, could mediate hippocampal-neuron survival (By similarity).

(Microbial infection) Acts as a coreceptor (CD4 being the primary receptor) for human immunodeficiency virus-1/HIV-1 X4 isolates and as a primary receptor for some HIV-2 isolates. Promotes Env-mediated fusion of the virus.

PTMs:

Phosphorylated on agonist stimulation. Rapidly phosphorylated on serine and threonine residues in the C-terminal. Phosphorylation at Ser-324 and Ser-325 leads to recruitment of ITCH, ubiquitination and protein degradation.

Ubiquitinated after ligand binding, leading to its degradation. Ubiquitinated by ITCH at the cell membrane on agonist stimulation. The ubiquitin-dependent mechanism, endosomal sorting complex required for transport (ESCRT), then targets CXCR4 for lysosomal degradation. This process is dependent also on prior Ser-/Thr-phosphorylation in the C-terminal of CXCR4. Also binding of ARRB1 to STAM negatively regulates CXCR4 sorting to lysosomes though modulating ubiquitination of SFR5S.

Sulfation on Tyr-21 is required for efficient binding of CXCL12/SDF-1alpha and promotes its dimerization. Tyr-7 and Tyr-12 are sulfated in a sequential manner after Tyr-21 is almost fully sulfated, with the binding affinity for CXCL12/SDF-1alpha increasing with the number of sulfotyrosines present. Sulfotyrosines Tyr-7 and Tyr-12 occupy clefts on opposing CXCL12 subunits, thus bridging the CXCL12 dimer interface and promoting CXCL12 dimerization.

O- and N-glycosylated. Asn-11 is the principal site of N-glycosylation. There appears to be very little or no glycosylation on Asn-176. N-glycosylation masks coreceptor function in both X4 and R5 laboratory-adapted and primary HIV-1 strains through inhibiting interaction with their Env glycoproteins. The O-glycosylation chondroitin sulfate attachment does not affect interaction with CXCL12/SDF-1alpha nor its coreceptor activity.

Subcellular Location:

Cell membrane>Multi-pass membrane protein. Cell junction. Early endosome. Late endosome. Lysosome.
Note: In unstimulated cells, diffuse pattern on plasma membrane. On agonist stimulation, colocalizes with ITCH at the plasma membrane where it becomes ubiquitinated. In the presence of antigen, distributes to the immunological synapse forming at the T-cell-APC contact area, where it localizes at the peripheral and distal supramolecular activation cluster (SMAC).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed in numerous tissues, such as peripheral blood leukocytes, spleen, thymus, spinal cord, heart, placenta, lung, liver, skeletal muscle, kidney, pancreas, cerebellum, cerebral cortex and medulla (in microglia as well as in astrocytes), brain microvascular, coronary artery and umbilical cord endothelial cells. Isoform 1 is predominant in all tissues tested.

Subunit Structure:

Monomer. Can form homodimers. Interacts with CD164. Interacts with ARRB2; the interaction is dependent on the C-terminal phosphorylation of CXCR4 and allows activation of MAPK1 and MAPK3. Interacts with ARRC; the interaction is dependent on the C-terminal phosphorylation of CXCR4 and modulates calcium mobilization. Interacts with RNF113A; the interaction, enhanced by CXCL12, promotes CXCR4 ubiquitination and subsequent degradation. Interacts (via the cytoplasmic C-terminal) with ITCH (via the WW domains I and II); the interaction, enhanced by CXCL12, promotes CXCR4 ubiquitination and leads to its degradation. Interacts with extracellular ubiquitin. Interacts with DBN1; this interaction is enhanced by antigenic stimulation. Following LPS binding, may form a complex with GDF5, HSP90AA1 and HSPA8.

(Microbial infection) Interacts with HIV-1 surface protein gp120 and Tat.

(Microbial infection) Interacts with HHV-8 protein ORF K4.

(Microbial infection) May interact with human cytomegalovirus/HHV-5 protein UL78.

(Microbial infection) Interacts with Staphylococcus aureus protein SSL10.

Family&Domains:

The amino-terminus is critical for ligand binding. Residues in all four extracellular regions contribute to HIV-1 coreceptor activity.

Belongs to the G-protein coupled receptor 1 family.

Research Fields

· Cellular Processes > Transport and catabolism > Endocytosis.   (View pathway)

· Environmental Information Processing > Signaling molecules and interaction > Cytokine-cytokine receptor interaction.   (View pathway)

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Organismal Systems > Immune system > Chemokine signaling pathway.   (View pathway)

· Organismal Systems > Development > Axon guidance.   (View pathway)

· Organismal Systems > Immune system > Leukocyte transendothelial migration.   (View pathway)

· Organismal Systems > Immune system > Intestinal immune network for IgA production.   (View pathway)

Restrictive clause

 

Affinity Biosciences tests all products strictly. Citations are provided as a resource for additional applications that have not been validated by Affinity Biosciences. Please choose the appropriate format for each application and consult Materials and Methods sections for additional details about the use of any product in these publications.

For Research Use Only.
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