cxcl12 2716 sd ligands (R&D Systems)

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R&D Systems cxcl12 2716 sd ligands
$Figure 3 CXCR7 expression and localization are modulated by CXCL11 and <t>CXCL12.</t> (A) Immunofluorescence staining of CXCR7 in AD-LNCaP cells treated with vehicle (0.1% BSA), CXCL11 (100 nM), or CXCL12 (100 nM) for 30 min. Nuclei and F-actin are labeled with DAPI and Texas-red phalloidin, respectively. (B) Western blot of cytosolic, membrane, and nuclear protein fractions isolated from LNCaP cells, cultured as described in (A) with pAbCXCR7 antibody. (C) Immunofluorescence staining of CXCR4 in AD-LNCaP cells as described in (A). (D-E) Western blot of cytosolic, membrane, and nuclear protein fractions isolated from LNCaP cells, cultured as described in (A), with antibodies to (D) CXCR4, and (E) AR, GM130, and histone H3. Silver staining demonstrates equivalent loading across samples. The densitometry values were normalized to BSA-treated samples for each subcellular compartment and labeled below the blots.$
Cxcl12 2716 Sd Ligands, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cxcl12+2716+sd+ligands/Recombinant+Human%2FFeline+CXCL12%2FSDF-1+beta+(aa+19-93)/pm25884570-61-22-25
Average 90 stars, based on 3 article reviews

cxcl12 2716 sd ligands - by Bioz Stars, 2026-10

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1) Product Images from "Androgen receptor and chemokine receptors 4 and 7 form a signaling axis to regulate CXCL12-dependent cellular motility."

Article Title: Androgen receptor and chemokine receptors 4 and 7 form a signaling axis to regulate CXCL12-dependent cellular motility.

Journal: BMC cancer

doi: 10.1186/s12885-015-1201-5

$Figure 3 CXCR7 expression and localization are modulated by CXCL11 and CXCL12. (A) Immunofluorescence staining of CXCR7 in AD-LNCaP cells treated with vehicle (0.1% BSA), CXCL11 (100 nM), or CXCL12 (100 nM) for 30 min. Nuclei and F-actin are labeled with DAPI and Texas-red phalloidin, respectively. (B) Western blot of cytosolic, membrane, and nuclear protein fractions isolated from LNCaP cells, cultured as described in (A) with pAbCXCR7 antibody. (C) Immunofluorescence staining of CXCR4 in AD-LNCaP cells as described in (A). (D-E) Western blot of cytosolic, membrane, and nuclear protein fractions isolated from LNCaP cells, cultured as described in (A), with antibodies to (D) CXCR4, and (E) AR, GM130, and histone H3. Silver staining demonstrates equivalent loading across samples. The densitometry values were normalized to BSA-treated samples for each subcellular compartment and labeled below the blots.$
Figure Legend Snippet: Figure 3 CXCR7 expression and localization are modulated by CXCL11 and CXCL12. (A) Immunofluorescence staining of CXCR7 in AD-LNCaP cells treated with vehicle (0.1% BSA), CXCL11 (100 nM), or CXCL12 (100 nM) for 30 min. Nuclei and F-actin are labeled with DAPI and Texas-red phalloidin, respectively. (B) Western blot of cytosolic, membrane, and nuclear protein fractions isolated from LNCaP cells, cultured as described in (A) with pAbCXCR7 antibody. (C) Immunofluorescence staining of CXCR4 in AD-LNCaP cells as described in (A). (D-E) Western blot of cytosolic, membrane, and nuclear protein fractions isolated from LNCaP cells, cultured as described in (A), with antibodies to (D) CXCR4, and (E) AR, GM130, and histone H3. Silver staining demonstrates equivalent loading across samples. The densitometry values were normalized to BSA-treated samples for each subcellular compartment and labeled below the blots.

Techniques Used: Expressing, Immunofluorescence, Staining, Labeling, Western Blot, Membrane, Isolation, Cell Culture, Silver Staining

Figure 6 CXCR7 modulates AR transcriptional activity. (A) Luciferase assay testing the effects of CXCR7 overexpression on the AR-target promoter probasin in LNCaP cells. LNCaP cells were co-transfected with the pGL4.10-Luc2-probasin and pRLSV40 Renilla vectors along with increasing amounts (30 ng experimental + 270 ng pcDNA3, 100 ng experimental + 200 ng pcDNA3, 300 ng experimental + 0 ng pcDNA3) of CXCR7 or ACTN4 cDNA mammalian expression vectors. The maximal amount (300 ng) of the pcDNA3 mammalian expression vector served as the positive control. Cells were subsequently treated with androgen (1 nM R1881) or vehicle (ethanol) and tested for dual luciferase activity. Student’s t-test was used to calculate significant differences (*p ≤0.05, n = 3) between control and experimental cells within the androgen-treatment group. (B) Luciferase assay testing the effects of CXCR7 siRNA knockdown and treatment with CXCR7 ligand on the AR-target promoter probasin. LNCaP cells were co-transfected with the pGL4.10-Luc2-probasin and pRLSV40-renilla vectors, along with control or experimental siRNAs (50 nM). Next, cells were pre-treated with indicated ligands (BSA, CXCL11, or CXCL12) for 30 min. Cells were then subsequently treated with androgen (1 nM R1881) or vehicle (ethanol) for 18 hrs and tested for dual luciferase activity. Student’s t-test was used to calculate significant differences (*p ≤0.05, n = 3) between control cells and experimental cells within the androgen-treatment group. (C) RNA isolated from LNCaP cells treated with vehicle (0.1% BSA), CXCL11 (10 nM), or CXCL12 (10 nM) for 30 min and subsequently treated with vehicle or androgen (1 nM R1881) for 18 hrs were subjected to qPCR analysis for AR, FASN, NKX3.1, PSA, and TMPRSS2 gene expressions. Student’s t-test was used to calculate significant differences (*p ≤0.05, n = 3) between control and chemokine ligand-treated cells.

Figure Legend Snippet: Figure 6 CXCR7 modulates AR transcriptional activity. (A) Luciferase assay testing the effects of CXCR7 overexpression on the AR-target promoter probasin in LNCaP cells. LNCaP cells were co-transfected with the pGL4.10-Luc2-probasin and pRLSV40 Renilla vectors along with increasing amounts (30 ng experimental + 270 ng pcDNA3, 100 ng experimental + 200 ng pcDNA3, 300 ng experimental + 0 ng pcDNA3) of CXCR7 or ACTN4 cDNA mammalian expression vectors. The maximal amount (300 ng) of the pcDNA3 mammalian expression vector served as the positive control. Cells were subsequently treated with androgen (1 nM R1881) or vehicle (ethanol) and tested for dual luciferase activity. Student’s t-test was used to calculate significant differences (*p ≤0.05, n = 3) between control and experimental cells within the androgen-treatment group. (B) Luciferase assay testing the effects of CXCR7 siRNA knockdown and treatment with CXCR7 ligand on the AR-target promoter probasin. LNCaP cells were co-transfected with the pGL4.10-Luc2-probasin and pRLSV40-renilla vectors, along with control or experimental siRNAs (50 nM). Next, cells were pre-treated with indicated ligands (BSA, CXCL11, or CXCL12) for 30 min. Cells were then subsequently treated with androgen (1 nM R1881) or vehicle (ethanol) for 18 hrs and tested for dual luciferase activity. Student’s t-test was used to calculate significant differences (*p ≤0.05, n = 3) between control cells and experimental cells within the androgen-treatment group. (C) RNA isolated from LNCaP cells treated with vehicle (0.1% BSA), CXCL11 (10 nM), or CXCL12 (10 nM) for 30 min and subsequently treated with vehicle or androgen (1 nM R1881) for 18 hrs were subjected to qPCR analysis for AR, FASN, NKX3.1, PSA, and TMPRSS2 gene expressions. Student’s t-test was used to calculate significant differences (*p ≤0.05, n = 3) between control and chemokine ligand-treated cells.

Techniques Used: Activity Assay, Luciferase, Over Expression, Transfection, Expressing, Plasmid Preparation, Positive Control, Control, Knockdown, Isolation

Figure 8 CXCR7 knockdown leads to a reduction in CXCR4 protein levels in LNCaP cells. (A) Transwell assay assessing the effects of CXCL12 on LNCaP migration. ANOVA was used to determine significant differences between vehicle (0.1% BSA) and CXCL12-treated cells (*p ≤0.05, n = 3). (B) Transwell assay assessing the effects of CXCR4 and CXCR7 knockdown on CXCL12-induced LNCaP cell migration. LNCaP cells transfected with 100 nM scrambled control, CXCR4, or CXCR7 siRNAs were seeded to the top chamber of the insert. The bottom chamber contained medium with 1% CS serum with 1 nM R1881 and CXCL12 at 0, 0.003, 0.03, or 0.3 nM concentrations. Data was normalized to control siRNA transfected, vehicle-treated cells. ANOVA was used to determine significant differences (*p ≤0.05, n = 3) between control and experimental cells. (C-E) Western blots to test the effects of CXCR7 and CXCR4 knockdown on AR signaling in LNCaP cells. Cells were transfected with control, CXCR7 or CXCR4 siRNA for 72 hrs and probed with antibodies against (C) CXCR7, (D) CXCR4, (E) AR, and PSA. The densitometry values were normalized to control siRNA transfected cells and labeled below the blots. (F) Silver staining demonstrates equivalent loading across the samples.

Figure Legend Snippet: Figure 8 CXCR7 knockdown leads to a reduction in CXCR4 protein levels in LNCaP cells. (A) Transwell assay assessing the effects of CXCL12 on LNCaP migration. ANOVA was used to determine significant differences between vehicle (0.1% BSA) and CXCL12-treated cells (*p ≤0.05, n = 3). (B) Transwell assay assessing the effects of CXCR4 and CXCR7 knockdown on CXCL12-induced LNCaP cell migration. LNCaP cells transfected with 100 nM scrambled control, CXCR4, or CXCR7 siRNAs were seeded to the top chamber of the insert. The bottom chamber contained medium with 1% CS serum with 1 nM R1881 and CXCL12 at 0, 0.003, 0.03, or 0.3 nM concentrations. Data was normalized to control siRNA transfected, vehicle-treated cells. ANOVA was used to determine significant differences (*p ≤0.05, n = 3) between control and experimental cells. (C-E) Western blots to test the effects of CXCR7 and CXCR4 knockdown on AR signaling in LNCaP cells. Cells were transfected with control, CXCR7 or CXCR4 siRNA for 72 hrs and probed with antibodies against (C) CXCR7, (D) CXCR4, (E) AR, and PSA. The densitometry values were normalized to control siRNA transfected cells and labeled below the blots. (F) Silver staining demonstrates equivalent loading across the samples.

Techniques Used: Knockdown, Transwell Assay, Migration, Transfection, Control, Western Blot, Labeling, Silver Staining

Control:

Article Title: Androgen receptor and chemokine receptors 4 and 7 form a signaling axis to regulate CXCL12-dependent cellular motility.
Article Snippet: .. The following reagents were purchased from the indicated vendors: AR agonist R1881 (methyltrienolone) (Perkin Elmer Life Sciences, Waltham, MA); CXCL11 (672-IT) and CXCL12 (2716-SD) ligands (R&D Systems, Minneapolis, MN); double-stranded experimentally validated siRNAs for scrambled control (1027281), AR (SI02757258), CXCR4 (SI02664235), CXCR7 (SI02660644) (Qiagen, Valencia, CA), and CXCR7 (109229) (Life Technologies, Chicago, IL); RNeasy Mini kit, RT2 qPCR primers for AR (PPH01016A), CXCR7 (PPH01182F), CXCR4 (PPH0 0621A), PSA (PPH01002B), FASN (PPH01012B), NK X3.1 (PPH02267C), TMPRSS2 (PPH02262C) (Qiagen); Oligofectamine Transfection Reagent, 4%-12% SDSpolyacrylamide gels, Superscript III enzyme, CyQUANT Cell Proliferation Assay Kit (Life Technologies); iQ SYBR-Green Supermix, Precision Plus Prestained Protein Standards, goat anti-mouse horseradish peroxidase (HRP)-conjugated secondary antibody, goat anti-rabbit HRP-conjugated secondary antibody (BioRad, Hercules, CA); mouse monoclonal AR antibody (AR441), rabbit polyclonal AR antibody (N-20), mouse monoclonal SBP antibody (SB19-C4) (Santa Cruz Biotechnology, Santa Cruz, CA); rabbit polyclonal CXCR7 antibodies (ab38089 [a.a. 1–100], and ab72100 [a.a. 106–117, QHNQWPM GELTC]), rabbit polyclonal CXCR4 antibody (ab2074) (Abcam, Cambridge, MA); rabbit polyclonal CXCR4 antibody (PAB9849) (Abnova, Taipei, Taiwan); mouse monoclonal GM130 antibody (BD Transduction Laboratories, San Jose, CA); rabbit polyclonal PSA antibody (DAKO, Carpinteria, CA); mouse monoclonal PSMA antibody (Meridian Life Science Inc, Memphis, TN); rabbit polyclonal Histone H3 antibody, rabbit monoclonal GAPDH antibody (14C10) (Cell Signaling Technology, Beverly, MA); BCA Protein Assay Kit and ECL Western Blotting Substrate Kit (ThermoFisher Scientific, Waltham, MA); Hyperfilm ECL film (GE Healthcare, Piscataway, NJ); fetal bovine serum, charcoal-stripped fetal bovine serum (Hyclone Laboratories, Logan, UT); GeneRuler 1 kb DNA Ladder (MBI Fermentas, Hanover, MD); Protein Deglycosylation Mix (P6039S, New England BioLabs, Ipswich, MA); Synthetic peptides to CXCR7 (a.a. 348–362, RVSET EYSALEQSTK) and AR (a.a. 299–315, KSTEDTAEYS PFKGGY) were synthesized by Alpha Diagonistics (San Antonio, TX). .. Generation of the polyclonal rabbit pAbCXCR7 antibody to CXCR7 The pAbCXCR7 polyclonal antibody was generated, by Alpha Diagnostics, against a synthetic peptide encoded by NCBI’s original CXCR7 cDNA sequence, which contained asparagine and alanine residues at amino acid positions 360 and 361 (348-RVSETEYSALEQNAK-362).

Article Title: Androgen receptor and chemokine receptors 4 and 7 form a signaling axis to regulate CXCL12-dependent cellular motility
Article Snippet: .. The following reagents were purchased from the indicated vendors: AR agonist R1881 (methyltrienolone) (Perkin Elmer Life Sciences, Waltham, MA); CXCL11 (672-IT) and CXCL12 (2716-SD) ligands (R&D Systems, Minneapolis, MN); double-stranded experimentally validated siRNAs for scrambled control (1027281), AR (SI02757258), CXCR4 (SI02664235), CXCR7 (SI02660644) (Qiagen, Valencia, CA), and CXCR7 (109229) (Life Technologies, Chicago, IL); RNeasy Mini kit, RT 2 qPCR primers for AR (PPH01016A), CXCR7 (PPH01182F), CXCR4 (PPH00621A), PSA (PPH01002B), FASN (PPH01012B), NKX3.1 (PPH02267C), TMPRSS2 (PPH02262C) (Qiagen); Oligofectamine Transfection Reagent, 4%-12% SDS-polyacrylamide gels, Superscript III enzyme, CyQUANT Cell Proliferation Assay Kit (Life Technologies); iQ SYBR-Green Supermix, Precision Plus Prestained Protein Standards, goat anti-mouse horseradish peroxidase (HRP)-conjugated secondary antibody, goat anti-rabbit HRP-conjugated secondary antibody (BioRad, Hercules, CA); mouse monoclonal AR antibody (AR441), rabbit polyclonal AR antibody (N-20), mouse monoclonal SBP antibody (SB19-C4) (Santa Cruz Biotechnology, Santa Cruz, CA); rabbit polyclonal CXCR7 antibodies (ab38089 [a.a. 1–100], and ab72100 [a.a. 106–117, QHNQWPMGELTC]), rabbit polyclonal CXCR4 antibody (ab2074) (Abcam, Cambridge, MA); rabbit polyclonal CXCR4 antibody (PAB9849) (Abnova, Taipei, Taiwan); mouse monoclonal GM130 antibody (BD Transduction Laboratories, San Jose, CA); rabbit polyclonal PSA antibody (DAKO, Carpinteria, CA); mouse monoclonal PSMA antibody (Meridian Life Science Inc, Memphis, TN); rabbit polyclonal Histone H3 antibody, rabbit monoclonal GAPDH antibody (14C10) (Cell Signaling Technology, Beverly, MA); BCA Protein Assay Kit and ECL Western Blotting Substrate Kit (ThermoFisher Scientific, Waltham, MA); Hyperfilm ECL film (GE Healthcare, Piscataway, NJ); fetal bovine serum, charcoal-stripped fetal bovine serum (Hyclone Laboratories, Logan, UT); GeneRuler 1 kb DNA Ladder (MBI Fermentas, Hanover, MD); Protein Deglycosylation Mix (P6039S, New England BioLabs, Ipswich, MA); Synthetic peptides to CXCR7 (a.a. 348–362, RVSETEYSALEQSTK) and AR (a.a. 299–315, KSTEDTAEYSPFKGGY) were synthesized by Alpha Diagonistics (San Antonio, TX). .. The pAbCXCR7 polyclonal antibody was generated, by Alpha Diagnostics, against a synthetic peptide encoded by NCBI’s original CXCR7 cDNA sequence, which contained asparagine and alanine residues at amino acid positions 360 and 361 (348-RVSETEYSALEQ NA K-362).

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cxcl12 2716 sd ligands (R&D Systems)

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