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Products:Immunology >> Innate Immunity >> Chemokines >> Alpha Chemokine Rec. (CXCR)
Overview
Product name
Anti-CXCR4 antibody
See all CXCR4 products (19) ...
Description
Rabbit polyclonal to CXCR4
Tested applications
Flow Cyt, IHC-P, IHC-Fr, IP, WB, ICC/IFmore details
Cross reactivity
Reacts with
Mouse, Rat, Human
Immunogen
Synthetic peptide, corresponding to N terminal amino acids 1-14 of Human CXCR4(Peptide available as ab8126.)
Epitope
MEGISIYTSDNYTE
Positive control
HeLa whole cell lysate.
General notes
At least 2 isoforms are known to exist.
Properties
Form
Liquid
Storage instructions
Store at +4°C.
Storage buffer
PBS with 0.02% sodium azide
Concentration
Concentration information loading...
Purity
IgG fraction
Primary antibody notes
HIV and chemokine receptor.
Clonality
Polyclonal
Isotype
IgG
Research areas
Immunology >> Immune System Diseases >> Antiviral Signaling >> HIV-related
Cancer >> Cancer Metabolism >> Response to hypoxia
Stem Cells >> Endothelial Progenitors >> Endothelial Markers
Immunology >> Adaptive Immunity >> Regulatory T Cells
Stem Cells >> Hematopoietic Progenitors >> Surface Molecules
Stem Cells >> Neural Stem Cells >> Surface Molecules
Neuroscience >> Neurology process >> Growth and Development >> Axonal Guidance Proteins
Microbiology >> Interspecies Interaction >> Host Virus Interaction
Immunology >> Innate Immunity >> Chemokines >> Alpha Chemokine Rec. (CXCR)
Applications
Show applications key
Our Abpromise guarantee covers the use of ab2074 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Flow Cyt: 1/50 - 1/100.
IHC-P: 1/50
IHC-Fr: 1/2000(Fix with 4% paraformaldehyde. Permeabilise with methanol. Amplification necessary.)
IP: Use a concentration of 10 µg/ml
WB: Use a concentration of 0.5 - 2 µg/ml.Detects a band of approximately 43 kDa (predicted molecular weight: 39 kDa).(To block use 5% non-fat dry milk in TBS to block for 2 hours at RT or overnight at 4°C.)
ICC/IF: 1/100
Target
Function
Receptor for the C-X-C chemokine CXCL12/SDF-1 that transduces a signal by increasing intracellular calcium ions levels and enhancing MAPK1/MAPK3 activation. Acts as a receptor for extracellular ubiquitin; leading to enhance intracellular calcium ions and reduce cellular cAMP levels. Involved in haematopoiesis and in cardiac ventricular septum formation. Plays also an essential role in vascularization of the gastrointestinal tract, probably by regulating vascular branching and/or remodeling processes in endothelial cells. Could be involved in cerebellar development. In the CNS, could mediate hippocampal-neuron survival. Acts as a coreceptor (CD4 being the primary receptor) for HIV-1 X4 isolates and as a primary receptor for some HIV-2 isolates. Promotes Env-mediated fusion of the virus.
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.
Involvement in disease
Defects in CXCR4 are a cause of WHIM syndrome (WHIM) [MIM:193670]; also known as warts, hypogammaglobulinemia, infections and myelokathexis. WHIM syndrome is an immunodeficiency disease characterized by neutropenia, hypogammaglobulinemia and extensive human papillomavirus (HPV) infection. Despite the peripheral neutropenia, bone marrow aspirates from affected individuals contain abundant mature myeloid cells, a condition termed myelokathexis.
Sequence similarities
Belongs to the G-protein coupled receptor 1 family.
Domain
The amino-terminus is critical for ligand binding. Residues in all four extracellular regions contribute to HIV-1 coreceptor activity.
Post-translational
modifications
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 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.
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.
Cellular localization
Cell membrane. In unstimulated cells, diffuse pattern on plasma membrane. On agonist stimulation, colocalizes with ITCH at the plasma membrane where it becomes ubiquitinated.
Target information above from: UniProt accessionP61073
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- FB22 antibody
- C-X-C chemokine receptor type 4 antibody
- CD184 antibody
see all
Database links
- Entrez Gene: 7852 Human
- Entrez Gene: 12767 Mouse
- Entrez Gene: 60628 Rat
- Omim: 162643 Human
- SwissProt: P61073 Human
- SwissProt: P70658 Mouse
- SwissProt: O08565 Rat
- Unigene: 593413 Human
see all
Anti-CXCR4 antibody images:
Western blot - CXCR4 antibody (ab2074)
Anti-CXCR4 antibody (ab2074) at 1/2000 dilution + HeLa whole cell lysate
Predicted band size : 39 kDa
Immunocytochemistry - CXCR4 antibody (ab2074)
Immunocytochemistry of CXCR4 in HeLa cells with CXCR4 antibody ab2074 at 2 µg/ml.
Immunocytochemistry/ Immunofluorescence - CXCR4 antibody (ab2074)
ab2074 at 1/100 staining differentiated rat skeletal myocytes by ICC/IF. The cells were 2% paraformaldehyde fixed for 15 minutes and then incubated cells with ab2074 overnight at 4°C. The image demonstrates CXCR4 expressing skeletal myocytes (green-cytoplasmic and cell membrane localization) with DAPI (blue-nuclear counter stain) [Right panel] and DIC (phase) image of the same skeletal myocytes ([left panel].
This image is courtesy of an Abreview submitted by Dr Mal Niladri
Immunocytochemistry/ Immunofluorescence - CXCR4 antibody (ab2074)
ab2074 staining CXCR4 expressing cells (of mesenchymal origin) by ICC/IF. The cells were methanol fixed and blocked with BSA prior to incubation with the antibody for 1 hour. A FITC conjugated swine anti-rabbit antibody was used as the secondary. Nuclei were counterstained with DAPI.
This image is courtesy of an anonymous Abreview
Flow Cytometry - CXCR4 antibody (ab2074)
ab2074 staining cells from human B cell lymphoma by flow cytometry. Cells were cultured and prepared in FACS buffer. The primary antibody was diluted 1/50 and incubated with the sample for 20 minutes at 4°C. A FITC conjugated swine anti-rabbit antibody was used as the secondary.
This image is courtesy of an anonymous Abreview
Western blot - CXCR4 antibody (ab2074)
Lane 1 : Anti-CXCR4 antibody (ab2074) at 1/250 dilution
Lane 2 : Anti-CXCR4 antibody (ab2074) at 1/500 dilution
Lane 3 : Anti-CXCR4 antibody (ab2074) at 1/750 dilution
Lane 4 : Anti-CXCR4 antibody (ab2074) at 1/1000 dilution
Lane 5 : Anti-CXCR4 antibody (ab2074) at 1/1500 dilution
Lane 6 : Anti-CXCR4 antibody (ab2074) at 1/2000 dilution
Lane 1 : Lysate prepared from human Ewing`s sarcoma cells
Lane 2 : Lysate prepared from human Ewing`s sarcoma cells
Lane 3 : Lysate prepared from human Ewing`s sarcoma cells
Lane 4 : Lysate prepared from human Ewing`s sarcoma cells
Lane 5 : Lysate prepared from human Ewing`s sarcoma cells
Lane 6 : Lysate prepared from human Ewing`s sarcoma cells
Lysates/proteins at 500000 cells per lane.
Secondary
HRP-conjugated goat polyclonal to rabbit IgG at 1/4000 dilution
Performed under reducing conditions.
Predicted band size : 39 kDa
Observed band size : 43 kDa (why is the actual band size different from the predicted?)
Exposure time : 5 minutes
This image is a courtesy of Sabine Topka
Immunocytochemistry/ Immunofluorescence - CXCR4 antibody (ab2074)
ab2074 staining CXCR4 in rat bone marrow cells by Immunocytochemistry/ Immunofluorescence. The cells were formaldehyde fixed, permeabilised in 0.1% Triton X-100 and then blocked using 5% serum for 1 hour at 25°C. Samples were then incubated with primary antibody at 1/100 for 12 hours at 4°C. The secondary antibody used was a goat anti-rabbit IgG conjugated to Alexa Fluor® 488 (green) used at a 1/250 dilution. DAPI was used to stain the cell nuclei (blue).
Image courtesy of an anonymous Abreview
Immunohistochemistry (Frozen sections) - CXCR4 antibody (ab2074)
ab2074 staining CXCR4 in rat kidney tubular epithelium tissue by Immunohistochemistry (Frozen sections). The tissue was formaldehyde fixed and then blocked using 2% BSA for 2 hours at 25°C. Samples were then incubated with primary antibody at 1/1000 for 9 hours at 4°C. The secondary antibody used was a goat anti-rabbit IgG conjugated to Alexa Fluor® 488 (green) used at a 1/500 dilution. DAPI was used to stain the cell nuclei (blue).
Image courtesy of an anonymous Abreview.
Immunocytochemistry/ Immunofluorescence - CXCR4 antibody (ab2074)
ICC/IF image of ab2074 stained Hela cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2074, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
References for Anti-CXCR4 antibody (ab2074)
This product has been referenced in:
- Wynn RFet al. A small proportion of mesenchymal stem cells strongly express functionally active CXCR4 receptor capable of promoting migration to bone marrow. Blood : (2004).Read more (PubMed: 15251986) »
- Castellone MDet al. Functional expression of the CXCR4 chemokine receptor is induced by RET/PTC oncogenes and is a common event in human papillary thyroid carcinomas. Oncogene 23:5958-67 (2004).Read more (PubMed: 15184868) »
See all 34 publications for this product
Publishing research using ab2074? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"