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CXCR4 Antibody

CXCR4 Antibody

Item no.	PRS-1012-0.02mg
Manufacturer	ProSci
Amount	0.02 mg
Category	Neuroscience Neural Lineage Markers Neuroinflammation Metabolism Glucose Homeostasis Regulation of Appetite Molecular Targets for Neuroscience Aging & Neurological Disorders Antibodies Immunohistochemistry Western Blot ELISA Immunofluorescence Primary Antibodies
Type	Antibody Polyclonal
Format	Liquid
Applications	WB, IF, IHC, ELISA
Specific against	Human, Mouse, Rat
Host	Rabbit
ECLASS 10.1	32160702
ECLASS 11.0	32160702
UNSPSC	12352203
Alias	CXCR4 Antibody: FB22, HM89, LAP3, LCR1, NPYR, WHIM, CD184, LESTR, NPY3R, NPYRL, HSY3RR, NPYY3R, D2S201E
Available
Shipping	blue ice or RT
By Research Area	Chemokines & Cytokines, Stem Cell, Infectious Disease
Homology	Predicted species reactivity based on immunogen sequence: Pig: (100%), Bovine: (86%), Sheep: (86%)
Immunogen	Anti-CXCR4 antibody (1012) was raised against a peptide corresponding to 15 amino acids near the center of human CXCR4 isoform b. The immunogen is located within amino acids 170-220 of CXCR4.
Applications	WB: 1 - 2 μ g/mL; IHC-P: 5 μ g/mL; IF: 4 μ g/mL. Antibody validated: Western Blot in human, mouse, and rat samples; Immunohistochemistry and Immunofluorescence in human samples. All other applications and species not yet tested.
Positive Control 1	Cat. No. 1201 - HeLa Cell Lysate
Positive Control 2	Cat. No. 1202 - A431 Cell Lysate
Positive Control 3	Cat. No. 1210 - 293 Cell Lysate
Positive Control 4	Cat. No. 1211 - HepG2 Cell Lysate
Positive Control 5	Cat. No. 1282 - NIH/3T3 Cell Lysate
Positive Control 6	Cat. No. 1471 - Rat thymus Lysate
Predicted Molecular Weight	Predicted: 40kD Observed: 44kD (Post-modification: 2 N-linked glycosylation)
Validation	Independent Antibody Validation (Figure 2) shows similar CXCR4 expression profile in both human and mouse cell lines detected by two independent anti-CXCR4 antibodies that recognize different epitopes, 1009 against N-terminus ectodomain and 1012 against the second extracellular loop. CXCR4 proteins are detected in most of the cell lines, but not in THP-1 cells by the two independent antibodies. Additionally, Figure 2 shows the mouse CXCR4 protein in NIH/3T3 and C2C12 cell lines migrates slightly slower than human isoform b detected by both CXCR4 antibodies (1009 and 1012), which is well correlated with their calculated molecular masses (40.4 kDa vs 39.7 kDa). siRNA knockdown validation (Figure 3): Anti-CXCR4 antibody (1012) specificity was further verified by CXCR4 specific siRNA knockdown. CXCR4 signal in HeLa cells transfected with CXCR4 siRNAs was weaker in comparison with that in HeLa cells transfected with control siRNAs. Animal Species Reactivity (Figure 4): Anti-CXCR4 antibodies (1009 and 1012) can detect the expression of CXCR4 protein in rat thymus, but not in rat brain and rat heart.
Isoforms	Human CXCR4 has four isoforms, including isoform a (356aa, 40.2kD), isoform b (352aa, 39.7kD), isoform c (423aa, 47.6kD), and isoform d (385aa, 43.4kD). This antibody detects human isoform b only, but not other human isoforms. Mouse CXCR4 has two isoforms, including isoform 1 (359aa, 40.4kD) and isoform 2 (357aa, 40.2kD). Rat CXCR4 has only one isoform identified so far (349aa, 39.4kD).
Purification	CXCR4 Antibody is Antibody is affinity chromatography purified via peptide column.
Clonality	Polyclonal
Isotype	IgG
Conjugate	Unconjugated
Buffer	CXCR4 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration	1 mg/mL
Storage Conditions	CXCR4 antibody can be stored at 4˚ C for three months and -20˚ C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Related Products	Cat. No. 1012P – CXCR4 Peptide
Disclaimer	Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.
Modifications	None
Ncbi Official Symbol	CXCR4
Accession #	NP_003458
Protein Gi #	4503175
Ncbi Gene Id #	7852
User Note	Optimal dilutions for each application to be determined by the researcher.
Ncbi Official Symbol	CXCR4
Ncbi Official Full Name	chemokine (C-X-C motif) receptor 4
Ncbi Organism	Homo sapiens
Swissprot #	P61073
Background	CXCR4, a G-protein coupled receptor (GPCR) with seven transmembrane domains, is a CXC chemokine receptor specific for stromal-derived-factor-1 (SDF-1 or CXCL12). CXCR4 was initially discovered as one of the co-receptors for HIV entry into CD4+ T cells (1). Blocking CXCR4 could be potentially used as novel therapeutics for HIV treatment. CXCR4 signaling plays an important role in the migration, proliferation and quiescence of hematopoietic stem cell and their retention within the bone marrow, where it has high levels of SDF-1/CXCL12(2). It has been demonstrated that CXCR4 signaling mediates CD20 up-regulation on B cells (3). CXCR4 is highly expressed in more than 23 types of cancer, including breast cancer, ovarian cancer, melanoma, and prostate cancer, while there is very less or no expression of CXCR4 in healthy tissues. CXCR4 expression in cancer cells has been reported to be associated with tumor survival, growth and metastasis in tissues with high levels of SDF-1/CXCL12, such as lungs, liver and bone marrow (4, 5). CXCR4 has been shown to regulate neuronal migration, cell positioning and axon wiring (6, 7). CXCR4 mutant mice displayed aberrant neuronal distribution, which implicates the role in neuronal disorders such as epilepsy. CXCR4 is also involved in WHIM syndrome (8). WHIM mutations in CXCR4 were recently found in patients with Waldenstrom's macroglobulinemia, and these mutations are correlated to clinical resistance to ibrutinib (9, 10).
Background References 1	Dimitrov DS. Cell 1997; 91:721-730
Background References 2	Chen et al. Circ Res. 2010; 107:1083-93
Background References 3	Pavlasova et al. Blood 2016; 128: 1609-13
Background References 4	Sun et al. Cancer Metastasis Reviews 2010; 29:709-22
Background References 5	Balkwill F. Nature Reviews 2004; 4:540-50
Background References 6	Bagri et al. Development 2002; 129:4249-60
Background References 7	Yang et al. Development 2013; 140:4554-64
Background References 8	Balabanjian et al. The Journal of Clinical Investigation 2008; 118:1074-84
Background References 9	Hunter et al. Blood 2014; 123:1637-46
Background References 10	Treon et al. N. Engl. J. Med. 2015; 372:1430-40
Citation 1	Odemis et al. CXCR7 is an active component of SDF-1 signalling in astrocytes and Schwann cells. J Cell Sci. 2010; 123(Pt 7):1081-8. PMID: 20197403
Citation 2	Kozak et al. Segregation of CD4 and CXCR4 into distinct lipid microdomains in T lymphocytes suggests a mechanism for membrane destabilization by human immunodeficiency virus. J Virol. 2002; 76(4):1802-15.PMID: 11799176
Citation 3	Scala et al. Human melanoma metastases express functional CXCR4. Clin Cancer Res 2006; 12(8):2427-33. PMID: 16638848
Citation 4	Recasens-Zorzo et al. Pharmacological modulation of CXCR4 cooperates with BET bromodomain inhibition in diffuse large B-cell lymphoma.Haematologica. PMID: 29954928
Citation 5	Pandit et al. Chronic allergen challenge induces pulmonary extramedullary hematopoiesis. Exp Lung Res. 2011; 37(5):279-90. PMID: 21309736
Citation 6	Nobuko et al. microRNA-150 regulates mobilization and migration of bone marrow-derived mononuclear cells by targeting CXCR4. PLoS ONE 2011; 6(10):e23114. PMID: 22039399
Citation 7	Nimmagadda et al. Immunoimaging of CXCR4 expression in brain tumor xenografts using SPECT/CT. J Nucl Med. 2009; 50(7):1124-30.PMID: 19525448
Citation 8	Lee et al. Gene expression in temporal lobe epilepsy is consistent with increased release of glutamate by astrocytes. Mol Med. 2007; 13(1-2):1-13. PMID: 17515952
Citation 9	Terlika et al. Sustained exposure to nicotine leads to extramedullary hematopoiesis in the spleen. Stem Cells 2006; 24:2373-81. PMID: 16825610
Citation 10	Schabath et al. CD24 affects CXCR4 function in pre-B lymphocytes and breast carcinoma cells. J Cell Sci. 2006; 119(Pt 2):314-25. PMID: 16390867
1st Image Caption	Figure 1 Western Blot Validation of CXCR4 in HeLa Cells Loading: 15 μ g of lysates per lane.Antibodies: 1012 (1 μ g/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
2nd Image Caption	Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile Loading: 15 μ g of lysates per lane. Antibodies: 1009 (1 μ g/mL), 1012 (1 μ g/mL), and beta-actin (1 μ g/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
3rd Image Caption	Figure 3 Validation with CXCR4 siRNA Knockdown in HeLa Cells HeLa cells were transfected with control siRNAs (lane 1) or CXCR4 siRNAs (lane 2) Loading: 10 μ g of HeLa whole cell lysates per lane. Antibodies: 1012 (2 μ g/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
4th Image Caption	Figure 4 Animal Species Reactivity Loading: Lysates/proteins at 20 μ g per lane.Antibodies: 1009 (2 μ g/mL) or 1012 (2 μ g/mL). 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
5th Image Caption	Figure 5 Immunofluorescence Validation of CXCR4 Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa cells labeling CXCR4 with 1012 at 4 μ g/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red). Image showing both membrane and cytoplasmic staining on HeLa cell line.
6th Image Caption	Figure 6 Immunohistochemistry Validation of CXCR4 in Human Spleen Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-CXCR4 antibody (1012) at 5 μ g/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚ C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
7th Image Caption	Figure 7 KO Validation of CXCR4 by Flow Cytometry (Ödemis, et al., 2010) Astrocytes from wild-type or CXCR4 knockout mice were stained with primary antibodies against CXCR4 and FITC-labeled secondary antibodies, and subsequently subjected to flow cytometry. CXCR4−/− astrocytes (red) showed loss of CXCR4 cell-surface expression compared with wild-type cells (black).
8th Image Caption	Figure 8 Overexpression Validation of CXCR4 (Kozak et al., 2002) U87MG and U87MG-CXCR4 extracts were included as negative and positive controls, respectively, for CXCR4 detection with anti-CXCR4 antibodies.
9th Image Caption	Figure 9 WB Validation of CXCR4 in Human Metastatic Melanoma (Scala et al., 2006) CXCR4 protein was detected in the human metastatic melanoma cell lines and human melanoma cell line (colo38), but not in the human primary melanocytes (MPR1) with anti-CXCR4 antibodies.

Note: The presented information and documents (Manual, Product Datasheet, Safety Datasheet and Certificate of Analysis) correspond to our latest update and should serve for orientational purpose only. We do not guarantee the topicality. We would kindly ask you to make a request for specific requirements, if necessary.

All products are intended for research use only (RUO). Not for human, veterinary or therapeutic use.

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