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Cxcr4 antibody

Manufactured by Abcam
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Sourced in Germany, United States, China, United Kingdom

The CXCR4 antibody is a laboratory reagent used for the detection and analysis of the CXCR4 protein. CXCR4 is a chemokine receptor that plays a role in cell migration and signaling. The antibody can be used in various research applications, such as Western blotting, immunohistochemistry, and flow cytometry, to identify and study the CXCR4 protein.

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Lab products found in correlation

E cadherin, by Cell Signaling Technology (2 mentions) Goat anti rabbit igg antibody, by Abcam (1 mentions) Protease inhibitor, by Roche (1 mentions) Vimentin antibody, by Cell Signaling Technology (1 mentions) Cell lysis buffer, by Cell Signaling Technology (1 mentions) Ecl detection system, by Cytiva (1 mentions) β actin antibody, by Merck Group (1 mentions) P erk, by Cell Signaling Technology (1 mentions) P stat3, by Cell Signaling Technology (1 mentions) P akt, by Cell Signaling Technology (1 mentions) Chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions) Hrp conjugated affinipure goat anti rabbit igg, by Proteintech (1 mentions) Twist, by Proteintech (1 mentions) Smad2 3, by Cell Signaling Technology (1 mentions) Stat3, by Cell Signaling Technology (1 mentions) P jnk, by Cell Signaling Technology (1 mentions) β actin, by Proteintech (1 mentions) Cd133, by Proteintech (1 mentions) Nestin, by Proteintech (1 mentions) Vegfa, by Proteintech (1 mentions)

Close spelling variants of this product

CXCR4 (91 citations) Anti-CXCR4 antibody (23 citations) Rabbit anti-CXCR4 antibody (6 citations) Primary CXCR4 antibody (5 citations) Rabbit anti-CXCR4 primary antibody (5 citations) Anti-CXCR4 rabbit polyclonal antibody (4 citations) Anti-human CXCR4 antibody (3 citations) Rabbit anti-rat CXCR4 antibody (2 citations) APC)-conjugated anti- CXCR4 antibody (2 citations)

6 protocols using cxcr4 antibody

1

Immunofluorescence Assay for CXCR4 Expression

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The cells that were cultured on glass slides were fixed with formaldehyde and were permeabilized with 0.5% Triton X-100, then they were incubated with CXCR4 antibody and goat anti-rabbit IgG antibody (Abcam, Germany), respectively. Finally, the cells were counterstained with DAPI and observed under a fluorescent inverted microscope. The positive reaction can be seen as green, and the nucleus was blue.
Zheng J., Mao X., Wang D, & Xia S. (2022). Preconditioned MSCs Alleviate Cerebral Ischemia-Reperfusion Injury in Rats by Improving the Neurological Function and the Inhibition of Apoptosis. Brain Sciences, 12(5), 631.
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2

CXCR4 Protein Expression Analysis

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Proteins (30 ug) were extracted from whole cell lysates and separated on 10% SDS-PAGE gels as described previously [20 (link)]. The protein ribbons were then transferred to the PVDF membrane and closed with a PBS containing 5% BSA for 1 h. After incubation with CXCR4 antibody (1:2000, Abcam, Waltham, MA, USA) overnight at 4 °C, the protein band was incubated for 1 h with the corresponding secondary antibody (CST, 6883, Danvers, MA, USA). Finally, we visualized the protein bands using chemiluminescent reagents.
Jiang G., Zheng Z.Q., Zhang J., Tian Z., Li X., Yu Z., Wang Z., You W, & Chen G. (2023). Development and Validation of CXCR4 Nomogram-Based Immune Infiltration/Tumor Inflammation in Primary Glioblastoma. Brain Sciences, 13(8), 1152.
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3

Comprehensive Cell Protein Extraction

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The total cell extracts were obtained using a Cell Lysis Buffer (Cell Signaling Technology Inc., Danvers, MA) in the presence of protease inhibitor (Roche Applied Science, Indianapolis, IN) at 4°C for 30 min. Briefly, cells (5x106) were washed twice with cell wash solution. The extracted proteins (40 μg) was resolved by SDS-PAGE, transferred to nitrocellulose membranes using the Novex™ Bolt™ gel electrophoresis system and iBlot® transfer stack system (Life Technologies Corp., Grand Island, NY), and probed with the appropriate dilution of CXCR4 antibody (1:1000, ab2074, Abcam, Cambridge, MA), β-actin antibody (1:10000, A5441, Sigma, St. Louis, MO) Na, K-ATPase antibody (1:1000, #3010, Cell Signaling Technology Inc., Danvers, MA), E-Cadherin (1:1000, #3195, Cell Signaling, Danvers, MA), and Vimentin antibody (1:1000, #3932, Cell Signaling Technology Inc., Danvers, MA). Immunoreactivity was detected using an ECL detection system (GE Healthcare Bio-Sciences Corp., Pittsburgh, PA). Films were exposed at multiple time points to ensure that images were not saturated.
Lee H.H., Bellat V, & Law B. (2017). Chemotherapy induces adaptive drug resistance and metastatic potentials via phenotypic CXCR4-expressing cell state transition in ovarian cancer. PLoS ONE, 12(2), e0171044.
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4

Western Blot Analysis of Protein Expression

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The cells were harvested in radioimmunoprecipitation assay buffer and lysed on ice. From each sample, equal amounts of protein were employed for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to PVDF membranes. After being blocked with 5% nonfat milk/TBST, the membrane was successively incubated with the appropriate primary antibody and a secondary antibody. Every incubation was followed by 3 washes with TBST. Protein signals were determined using a chemiluminescent substrate (Thermo Fisher Scientific, Waltham, MA, USA). Primary antibodies: MSH6, N-cadherin, Vimentin, E-cadherin, p-AKT, p-STAT3, p-Smad2/3, p-ERK, p-p38, p-JNK, p-p65, STAT3, Smad2/3, ERK, p38, JNK and p65 antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA); Ki67, Nestin, CD133, SOX2, Cyclin D1, Bax, Bcl-2, MMP2, MMP9, HIF1A, VEGFA, Snail, Slug, Twist, ZEB1, ZEB2, AKT, TGFB1 and β-actin antibodies were supplied by Proteintech (Wuhan, Hubei, China); CXCR4 antibody was obtained from Abcam (Shanghai, China). Secondary antibodies: horseradish peroxidase (HRP)-conjugated affinipure goat anti-mouse IgG and HRP- conjugated affinipure goat anti-rabbit IgG were purchased from Proteintech (Wuhan, Hubei, China).
Chen Y., Liu P., Sun P., Jiang J., Zhu Y., Dong T., Cui Y., Tian Y., An T., Zhang J., Li Z, & Yang X. (2019). Oncogenic MSH6-CXCR4-TGFB1 Feedback Loop: A Novel Therapeutic Target of Photothermal Therapy in Glioblastoma Multiforme. Theranostics, 9(5), 1453-1473.
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5

Multiplex Immunofluorescence Staining of CXCR4 and gp100

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Immunofluorescence double staining with CXCR4 and gp100 was performed using Opal Automation Multiplex IHC Detection Kits (Akova Bioscience, Inc. Marlborough, MA) in Ventana DISCOVERY ULTRA System (Roche, Tucson, Arizona), according to the manufacturer’s instructions. Briefly, formalin-fixed paraffin-embedded tissue specimens were sliced into 5μM sections. Tissue sections were deparaffinized and blocked with a blocking buffer, then incubated with the CXCR4 antibody (Catalog number: ab124824; Abcam, Cambridge, MA, 1:400 dilution) for 40 min. Following a thorough wash, the tissue sections were incubated with the secondary antibody and developed with Opal520. Tissue sections were then incubated with anti-melanoma gp100 antibody (Catalog number: ab137078; Abcam, Cambridge, MA, 1:200 dilution) for 40 min. After a thorough wash, the tissue sections were incubated with the secondary antibody and developed with Opal570. DAPI was applied as the nuclear counterstaining. Images were scanned and analyzed with Phenochart TM 1.0 Whole Slide Contextual Viewer for Annotation and Review (Akova Bioscience, Inc. Marlborough, MA)
Yang H., Tan S., Qiao J., Xu Y., Gui Z., Meng Y., Dong B., Peng G., Ibhagui O.Y., Qian W., Lu J., Li Z., Wang G., Lai J., Yang L., Grossniklaus H.E, & Yang J.J. (2022). Non-invasive detection and complementary diagnosis of liver metastases via chemokine receptor 4 imaging. Cancer Gene Therapy, 29(12), 1827-1839.
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6

Quantification of PTX3 in Human Cells

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A human PTX3 Quantikine ELISA kit was purchased from R&D Systems (Minneapolis, MN, USA). Human anti-PTX3 antibody and CXCR4 antibody were obtained from Abcam (Cambridge, UK) and Thermo Fisher Scientific (Rockford, IL, USA), respectively. Human α-tubulin antibody was acquired from Biogenex (Fremont, CA, USA). Recombinant SDF-1α was purchased from PeproTech (Rocky Hill, NJ, USA). Methyl-thiazolyl-tetrazolium (MTT), β-glycerophosphate disodium salt pentahydrate, ascorbic acid, dexamethasone, and the alkaline phosphatase kit for ALP staining were all purchased from Sigma-Aldrich (St. Louis, MO, USA).
Kim Y., Park J.Y., Park H.J., Kim M.K., Kim Y.I., Kim H.J., Bae S.K, & Bae M.K. (2019). Pentraxin-3 Modulates Osteogenic/Odontogenic Differentiation and Migration of Human Dental Pulp Stem Cells. International Journal of Molecular Sciences, 20(22), 5778.
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