Recombinant human cxcl12

Manufactured by R&D Systems

Sourced in United States

Recombinant human CXCL12 is a chemokine protein produced in a recombinant system. CXCL12, also known as stromal cell-derived factor 1 (SDF-1), is a small cytokine that regulates the trafficking and homing of cells, particularly hematopoietic stem and progenitor cells.

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Lab products found in correlation

Fibronectin, by Merck Group (1 mentions) Ccl21, by R&D Systems (1 mentions) Ly294002, by Cell Signaling Technology (1 mentions) Pi3k p85 antibody, by Cell Signaling Technology (1 mentions) Phospho pi3k p85 tyr 458 p55 tyr199 antibody, by Cell Signaling Technology (1 mentions) Pten antibody, by Cell Signaling Technology (1 mentions) Anti human cxcl12 antibody, by R&D Systems (1 mentions) Akt antibody, by Cell Signaling Technology (1 mentions) Phospho akt ser473, by Cell Signaling Technology (1 mentions) Tc14012, by Merck Group (1 mentions) Calcein am, by Dojindo Laboratories (1 mentions) Amd3100, by Merck Group (1 mentions) Anti cxcl12 antibody, by R&D Systems (1 mentions) Fluo 4 am dye, by Thermo Fisher Scientific (1 mentions) Flipr tetra, by Molecular Devices (1 mentions) Adenosine 5 triphosphate, by Merck Group (1 mentions) Screenworks software, by Molecular Devices (1 mentions) Cellbind plate, by Corning (1 mentions) Dp80 ccd camera, by Olympus (1 mentions) Formaldehyde, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Recombinant human CXCL12 protein Recombinant Human/Rhesus Macaque/Feline CXCL12/SDF1α Recombinant Human/Rhesus Macaque/Feline CXCL12/SDF-1 alpha Recombinant human SDF-1α/CXCL12 Recombinant human CXCL12/SDF-1 Recombinant CXCL12 Human CXCL12 CXCL12

8 protocols using recombinant human cxcl12

Chemotaxis Assay for Immune Cell Migration

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Recombinant human CXCL12 and CCL21 were purchased from R&D Systems. A transwell cell migration assay was performed using a 24‐well chemotaxis chamber with 5‐µm pore inserts precoated with 20 µg/mL fibronectin (Sigma‐Aldrich) for 1 hour at room temperature. The cells (1 × 10⁵ cells/mL) were resuspended in DMEM/F12 supplemented with 2% FCS and added to the upper chambers, whereas CXCL12 or CCL21 to the lower chambers. After incubating for 18 hours, the cells on the lower surface of the membrane were fixed with methanol and stained with hematoxylin and eosin. Cell number was counted under a light microscope in eight fields per membrane at a 200‐fold magnification. The relative chemotactic index was determined as the ratio of the proportion of cells that migrated in response to multiple chemokines to the proportion that migrated in response to a single chemokine. The assay was performed in triplicate. Data were expressed as the mean ± SD and were analyzed by Student's t test. A two‐tailed P‐value of 0.05 or less was considered significant.

Hayasaka H., Yoshida J., Kuroda Y., Nishiguchi A., Matsusaki M., Kishimoto K., Nishimura H., Okada M., Shimomura Y., Kobayashi D., Shimazu Y., Taya Y., Akashi M, & Miyasaka M. (2022). CXCL12 promotes CCR7 ligand–mediated breast cancer cell invasion and migration toward lymphatic vessels. Cancer Science, 113(4), 1338-1351.

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CXCL12 Signaling Pathway Inhibition

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Recombinant Human CXCL12 and anti-human CXCL12 antibody were purchased by R&D system Inc. (Minneapolis, MN, USA). LY294002 (PI3K inhibitor) was ordered from Cell Signaling Technology (Beverly, MA, USA). The monoclonal antibodies (mAbs) included PTEN antibody, phospho-PTEN (ser380) antibody, Akt antibody, phospho-Akt (ser473), PI3K p85 antibody, phospho-PI3K p85 (Tyr 458) /p55 (Tyr199) antibody were provided by Cell Signaling Technology.

Ma J., Sun X., Wang Y., Chen B., Qian L, & Wang Y. (2019). Fibroblast-derived CXCL12 regulates PTEN expression and is associated with the proliferation and invasion of colon cancer cells via PI3k/Akt signaling. Cell Communication and Signaling : CCS, 17, 119.

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CXCL12 and CXCR4 Antibody Protocols

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Recombinant human CXCL12 was purchased from R&D Systems (Minneapolis, MN). Neutralizing monoclonal anti-human CXCL12 (anti-CXCL12 Ab), anti-human CXCR4 (anti-CXCR4 Ab) were obtained from Carbiochem (San Diego, CA, USA).

Ma J., Su H., Yu B., Guo T., Gong Z., Qi J., Zhao X, & Du J. (2018). CXCL12 gene silencing down-regulates metastatic potential via blockage of MAPK/PI3K/AP-1 signaling pathway in colon cancer. Clinical & Translational Oncology, 20(8), 1035-1045.

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Protein Adsorption on Microfluidic Devices

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Recombinant human CXCL12 and recombinant human Fc-ICAM1 chimeric fusion protein were obtained from R&D Systems (Minneapolis, MN). SDC4, expressed in mammalian cells so as to have glycosaminoglycans, was obtained from Adipogen Corporation (San Diego, CA). These molecules adsorbed onto the device walls at concentrations of 20, 80, and 50 micrograms per milliliter, for CXCL12, ICAM1, and SDC4, respectively, in phosphate buffered saline (PBS) for a minimum of four hours in a 37 °C incubator. After this incubation, device channels were blocked with 5% BSA in saline for a minimum of 30 minutes.

Benson B.L., Li L., Myers J.T., Dorand R.D., Gurkan U.A., Huang A.Y, & Ransohoff R.M. (2018). Biomimetic post-capillary venule expansions for leukocyte adhesion studies. Scientific Reports, 8, 9328.

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Chemoattractant CXCL12 and Receptor Assay

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Recombinant human CXCL12 was obtained from R&D Systems (Wiesbaden, Germany). Calcein-AM was purchased from Dojindo (Dojindo Laboratories, China). Polyclonal rabbit anti-CXCR4 and polyclonal rabbit anti-CXCR7 were obtained from Abcam (Cambridge, MA, USA). Polyclonal rabbit anti-β-Actin antibody, TC14012 and AMD3100, were obtained from Sigma (Deisenhofen, Germany). CCX771 was obtained from ChemoCentryx (Mountain View, CA, USA).

Zhang M., Qiu L., Zhang Y., Xu D., Zheng J.C, & Jiang L. (2017). CXCL12 enhances angiogenesis through CXCR7 activation in human umbilical vein endothelial cells. Scientific Reports, 7, 8289.

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CXCL12, IL-1α, and IL-1Ra Assays

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Recombinant human CXCL12 and anti-CXCL12 antibody were purchased from R&D systems (Minneapolis, MN), recombinant human IL-1α was provided by Diaclone (Beasancon, France), while recombinant human IL-1 Receptor Antagonist (IL-1Ra) was provided from Pepro Tech EC Ltd (London, UK).

Ma J., Liang W., Qiang Y., Li L., Du J., Pan C., Chen B., Zhang C., Chen Y, & Wang Q. (2021). Interleukin-1 receptor antagonist inhibits matastatic potential by down-regulating CXCL12/CXCR4 signaling axis in colorectal cancer. Cell Communication and Signaling : CCS, 19, 122.

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Measuring Ca2+ Dynamics in Breast Cancer Cells

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For Ca²⁺ measurements, MDA-MB-231 (7.5×10³ cells/well) or MDA-MB-468 (1.5 ×10⁴ cells/well) cells were seeded in a 96-well CellBIND plate (Corning Life Sciences, Corning, NY, USA) in antibiotic-free DMEM containing L-glutamine (4 mM) and 10% FBS (MDA-MB-468 cells were seeded at a higher density due to their slower proliferation rate). At 24 h post-plating, the FBS concentration was decreased to 8%. At 72 h post-plating, Ca²⁺ assays were performed using a fluorescence imaging plate reader, FLIPR^TETRA (Molecular Devices, LLC, Sunnyvale, CA, USA) and 4 µM Fluo-4 AM dye (Molecular Probes; Thermo Fisher Scientific, Inc.) in physiological salt solution, as previously described (24 (link)). Cells were excited at 470–495 nm and emission was assessed at 515–575 nm over an 800 sec period. Relative cytoplasmic (CYT) [Ca²⁺]_CYT was determined in the presence of 300 and 100 ng/ml recombinant human CXCL12 (R&D Systems, Inc., Minneapolis, MN, USA) or 100 µM adenosine 5′-triphosphate (ATP; Sigma-Aldrich; Merck KGaA). Data were acquired using ScreenWorks™ software (v2.0.0.27, Molecular Devices, LLC) and are presented as the response over baseline, which is a measure of relative [Ca²⁺]_CYT.

Jamaludin S.Y., Azimi I., Davis F.M., Peters A.A., Gonda T.J., Thompson E.W., Roberts-Thomson S.J, & Monteith G.R. (2018). Assessment of CXC ligand 12-mediated calcium signalling and its regulators in basal-like breast cancer cells. Oncology Letters, 15(4), 4289-4295.

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Quantifying α_V_β_3 Integrin Expression in CXCL12-Stimulated Cells

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C4-2B cells were grown in triplicate on glass coverslips in a six-well dish and then stimulated to express α_Vβ₃ by treatment with 200 ng/mL of recombinant human CXCL12 (R&D Systems, Minneapolis, MN) for 1, 6, or 12 h at 37 °C. After cells were stimulated, they were rinsed with PBS, fixed with 4% formaldehyde (Affymetrix) for 10 min at room temperature, washed twice with PBS, and blocked for 1 h with 1% BSA in PBS. Next cells were incubated for 4 h with a 1:100 dilution of anti-integrin α_Vβ₃ antibody, clone LM609 (Millipore). Incubation was followed by three washes with PBS and then incubation for 45 min with a 1:200 dilution of the goat anti-mouse IgG (H+L) secondary antibody, Alexa Fluor 488 conjugate (Life Technologies). Cells were washed twice more and mounted on slides with Prolong Gold antifade reagent with DAPI (Invitrogen). Fluorescent images were taken with Olympus BX53 fluorescence microscope with a DP80 CCD camera (Olympus). It is equipped with a 100 W halogen lamp and a condenser (NA = 0.9). The exposure time of 0.25 s and illumination intensity lamp voltage of 9 V were used for imaging. The images were analyzed by ImageJ (NIH) software.

Gdowski A.S., Lampe J.B., Lin V.J., Joshi R., Wang Y.C., Mukerjee A., Vishwanatha J.K, & Ranjan A.P. (2019). Bioinspired Nanoparticles Engineered for Enhanced Delivery to the Bone. ACS applied nano materials, 2(10), 6249-6257.

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