|
ArtNr |
PRS-1009-0.02mg |
|
Hersteller |
ProSci
|
|
Menge |
0.02 mg |
|
Kategorie |
|
|
Typ |
Antibody Polyclonal |
|
Format |
Liquid |
|
Applikationen |
WB, FC, IF, IP, ICC, ELISA, IHC-P |
|
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 |
|
Lieferbar |
 |
| Shipping |
blue ice or RT |
| By Research Area |
Chemokines & Cytokines, Stem Cell, Infectious Disease |
| Homology |
Predicted species reactivity based on immunogen sequence: Bovine: (86%), Pig: (71%) |
| Immunogen |
Anti-CXCR4 antibody (1009) was raised against a peptide corresponding to 14 amino acids near the amino terminus of human CXCR4 isoform b.
The immunogen is located within the first 50 amino acids of CXCR4. |
| Applications |
WB: 1 - 2 μ g/mL; IP/ ICC: 2 μ g/mL; IHC-P: 5 μ g/mL; IF: 20 μ g/mL; Flow Cyt: 0.1 μ g/mL.
Antibody validated: Western Blot in human, mouse, and rat samples; Immunohistochemistry, Immunocytochemistry and Immunofluorescence in human samples; Flow Cytometry in human and mouse samples. All other applications and species not yet tested. |
| Specificity |
CXCR4 Antibody is predicted to not cross-react with other CXCR familiy members. |
| 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: 40 kDa
Observed: 44 kDa (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 (1009) 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. Recombinant Protein Test (Figure 5) shows that anti-CXCR4 antibody (1009) detects partial recombinant CXCR4 protein that contains the amino acids of 1009 antigenic peptide. A band with same size can be observed in coomassie blue staining. |
| 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 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. 1009P – 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 |
| 1st Image Caption |
Figure 1 Western Blot Validation of CXCR4 in HeLa Cells
Loading: 15 μ g of lysates per lane. Antibodies: 1009 (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 in Cell Lines
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: 1009 (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 Recombinant Protein Test
Loading: CXCR4 partial recombinant protein (Novus Biologicals, Cat# H00007852-Q01). Lane 1: Anti-CXCR4 antibody (0.1 μ g/mL) 1 h incubation at RT in 5% NFDM/TBST. Lane 2: Coomassie blue staining. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. |
| 6th Image Caption |
Figure 6 Immunofluorescence Validation of CXCR4 in HeLa Cells
Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa cells labeling CXCR4 with 1009 at 20 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 cells. |
| 7th Image Caption |
Figure 7 Flow Cytometry Validation of CXCR4 in HeLa Cells
Overlay histogram showing HeLa cells stained with 1009 (red line, 1μ g/1x106 cells). 1 h incubation at 4˚ C in 2% FBS/PBS. Followed by secondary antibody 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 1 h 4˚ C.
Isotype control antibody (Green line) was mouse IgG1 (1μ g/1x106 cells) used under the same conditions. |
| 8th Image Caption |
Figure 8 Immunohistochemistry Validation of CXCR4 in Human Spleen
Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-CXCR4 antibody (1009) 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. |
| 9th Image Caption |
Figure 9 Immunocytochemistry Validation of CXCR4 in HeLa Cells
Immunocytochemical analysis of HeLa cells using anti-CXCR4 antibody (1009) at 2 μ g/ml. Cells 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. |
| 10th Image Caption |
Figure 10 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). |
| 11th Image Caption |
Figure 11 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. |
| 12th Image Caption |
Figure 12 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. |
Hinweis: Die dargestellten Informationen und Dokumente (Bedienungsanleitung, Produktdatenblatt, Sicherheitsdatenblatt und Analysezertifikat) entsprechen unserem letzten Update und sollten lediglich der Orientierung dienen. Wir übernehmen keine Garantie für die Aktualität. Für spezifische Anforderungen bitten wir Sie, uns eine Anfrage zu stellen.
Alle Produkte sind nur für Forschungszwecke bestimmt. Nicht für den menschlichen, tierärztlichen oder therapeutischen Gebrauch.
Mein Benutzerkonto
Preise & Rabatte einsehen
Geschenke sichern
Angebote anfragen & online annehmen
