CXCL12 Kit ELISA

N° du produit ABIN625084

Détail du produit CXCL12 Kit ELISA

Antigène: CXCL12 (Chemokine (C-X-C Motif) Ligand 12 (CXCL12))

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 80-6000 pg/mL

Seuil minimal de détection: 80 pg/mL

Application: ELISA

Fonction: Human SDF-1 alpha (CXCL12 alpha) ELISA Kit for cell culture supernatants, heparin, EDTA, or citrate treated plasma, and serum samples.

Type d'échantillon: Plasma, Cell Culture Supernatant, Serum

Analytical Method: Quantitative

Specificité: This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin, MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.

Sensibilité: < 80 pg/mL

Attributs du produit:

• Strip plates and additional reagents allow for use in multiple experiments
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of antibody array data

Ingrédients:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent(s)
• Lyophilized Standard
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Matériel non inclus:

• Distilled or deionized water
• Precision pipettes to deliver 2 μL to 1 μL volumes
• Adjustable 1-25 μL pipettes for reagent preparation
• 100 μL and 1 liter graduated cylinders
• Tubes to prepare standard and sample dilutions
• Absorbent paper
• Microplate reader capable of measuring absorbance at 450nm
• Log-log graph paper or computer and software for ELISA data analysis

Information d'application

Indications d'application: Recommended Dilution for serum and plasma samples2 fold

Volume d'échantillon: 100 μL

Plaque: Pre-coated

Protocole:

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 μL of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Préparation des réactifs:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
   2. Sample preparatuion: If your samples need to be diluted, Assay Diluent A (Item D) should be used for dilution of plasma samples. 1x Assay Diluent B (Item E) should be used for dilution of culture supernatants and urine. Suggested dilution for normal serum/plasma: 2 fold. Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator. Plasma: Collect plasma using heparin, EDTA or citrate as an anticoagulant. Centrifuge for 15 minutes at 1000 x g within 30 minutes of collection. An additional centrifugation step of the separated plasma at 10,000 x g for 10 minutes at 4° C is recommended for complete platelet removal (to measure circulating levels of SDF-1alpha, platelet- free plasma should be collected).
   3. Assay Diluent B should be diluted 5-fold with deionized or distilled water.
   4. Preparation of standard: Briefly spin the vial of Item C. Add 400 µL Assay Diluent A (for plasma samples) or 1x Assay Diluent B (for cell culture medium and urine) into Item C vial to prepare a 17 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 180 µL SDF-1alpha standard from the vial of Item C, into a tube with 330 µL Assay Diluent A or 1x Assay Diluent B to prepare a 6,000 pg/mL stock standard solution. Pipette 250 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Gently vortex to mix. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 pg/mL). 250 µL 180 µL standard + 330 µL 250myl 250 µL 250 µL 250 µL 250 µL 6,000 3,000 1,500 750 375 187.5 93.75 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
   5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
   6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent B into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure.
   7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 200-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 50 µL of HRP-Streptavidin concentrate into a tube with 10 ml 1x Assay Diluent B to prepare a 200-fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well

Procédure de l'essai:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
   2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
   3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
   4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
   5. Discard the solution. Repeat the wash as in step
   6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
   7. Discard the solution. Repeat the wash as in step
   8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
   9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.

Calcul des résultats:

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Assay Diluent A Human SDF-1-alpha concentration (pg/mL) 10 100 1000 10000 O D =4 50 n m 0.01 0.1 1 10 Assay Diluent B Human SDF-1-alpha concentration (pg/mL) 10 100 1000 10000 O D =4 50 n m 0.01 0.1 1 10
Sensitivity: The minimum detectable dose of SDF-1alpha is typically less than 80 pg/mL.
Recovery: Recovery was determined by spiking various levels of human SDF-1alpha into human plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Plasma 89.34 85-104 Cell culture media 94.63 86-104
Linearity: Sample Type Plasma Cell Culture Media 1:2 Average % of Expected 93 95 Range ( %) 83-102 84-104 1:4 Average % of Expected 94 96 Range ( %) 82-103 85-104
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

Précision du teste: Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %

Restrictions: For Research Use only

Stockage

Conseil sur la manipulation: Avoid repeated freeze-thaw cycles.

Stock: -20 °C

Stockage commentaire: The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Date de péremption: 6 months

Références pour CXCL12 Kit ELISA (ABIN625084)

Koob, Lim, Zabek, Massee: "Cytokines in single layer amnion allografts compared to multilayer amnion/chorion allografts for wound healing." dans: Journal of biomedical materials research. Part B, Applied biomaterials, Vol. 103, Issue 5, pp. 1133-40, (2015) (PubMed).

Liu, Weng, Yuan, Zhang, Bai, Li, Yang, Zhang, He, Yan, Zhan, Shi: "CXCL12/CXCR4 signal axis plays an important role in mediating bone morphogenetic protein 9-induced osteogenic differentiation of mesenchymal stem cells." dans: International journal of medical sciences, Vol. 10, Issue 9, pp. 1181-92, (2013) (PubMed).

Gravina, Mancini, Ranieri, Di Pasquale, Marampon, Di Clemente, Ricevuto, Festuccia: "Phenotypic characterization of human prostatic stromal cells in primary cultures derived from human tissue samples." dans: International journal of oncology, Vol. 42, Issue 6, pp. 2116-22, (2013) (PubMed).

Vizio, Biasi, Scirelli, Novarino, Prati, Ciuffreda, Montrucchio, Poli, Bellone: "Pancreatic-carcinoma-cell-derived pro-angiogenic factors can induce endothelial-cell differentiation of a subset of circulating CD34+ progenitors." dans: Journal of translational medicine, Vol. 11, pp. 314, (2013) (PubMed).

Dudás, Fullár, Romani, Pritz, Kovalszky, Hans Schartinger, Mathias Sprinzl, Riechelmann: "Curcumin targets fibroblast-tumor cell interactions in oral squamous cell carcinoma." dans: Experimental cell research, Vol. 319, Issue 6, pp. 800-9, (2013) (PubMed).

Shangguan, Ti, Krause, Hai, Zhao, Yang, Liu: "Inhibition of TGF-?/Smad signaling by BAMBI blocks differentiation of human mesenchymal stem cells to carcinoma-associated fibroblasts and abolishes their protumor effects." dans: Stem cells (Dayton, Ohio), Vol. 30, Issue 12, pp. 2810-9, (2012) (PubMed).

Sobrino, Pérez-Mato, Brea, Rodríguez-Yáñez, Blanco, Castillo: "Temporal profile of molecular signatures associated with circulating endothelial progenitor cells in human ischemic stroke." dans: Journal of neuroscience research, Vol. 90, Issue 9, pp. 1788-93, (2012) (PubMed).

Sobrino, Arias, Pérez-Mato, Agulla, Brea, Rodríguez-Yáñez, Castillo: "Cd34+ progenitor cells likely are involved in the good functional recovery after intracerebral hemorrhage in humans." dans: Journal of neuroscience research, Vol. 89, Issue 7, pp. 979-85, (2012) (PubMed).

Perez-Gracia, Prior, Guillén-Grima, Segura, Gonzalez, Panizo, Melero, Grande-Pulido, Gurpide, Gil-Bazo, Calvo: "Identification of TNF-alpha and MMP-9 as potential baseline predictive serum markers of sunitinib activity in patients with renal cell carcinoma using a human cytokine array." dans: British journal of cancer, Vol. 101, Issue 11, pp. 1876-83, (2009) (PubMed).

Sengenès, Miranville, Maumus, de Barros, Busse, Bouloumié: "Chemotaxis and differentiation of human adipose tissue CD34+/CD31- progenitor cells: role of stromal derived factor-1 released by adipose tissue capillary endothelial cells." dans: Stem cells (Dayton, Ohio), Vol. 25, Issue 9, pp. 2269-76, (2007) (PubMed).

Nagasawa, Hirota, Tachibana, Takakura, Nishikawa, Kitamura, Yoshida, Kikutani, Kishimoto: "Defects of B-cell lymphopoiesis and bone-marrow myelopoiesis in mice lacking the CXC chemokine PBSF/SDF-1." dans: Nature, Vol. 382, Issue 6592, pp. 635-8, (1996) (PubMed).

Détail du antigène

Antigène: CXCL12 (Chemokine (C-X-C Motif) Ligand 12 (CXCL12))

Autre désignation: SDF 1 alpha (CXCL12 Produits)

Synonymes: IRH Kit ELISA, PBSF Kit ELISA, SCYB12 Kit ELISA, SDF1 Kit ELISA, TLSF Kit ELISA, TPAR1 Kit ELISA, CINC-1 Kit ELISA, Gro1 Kit ELISA, Pbsf Kit ELISA, Scyb12 Kit ELISA, Sdf1 Kit ELISA, Tlsf Kit ELISA, Tpar1 Kit ELISA, Fsp Kit ELISA, KC Kit ELISA, Mgsa Kit ELISA, N51 Kit ELISA, Scyb1 Kit ELISA, gro Kit ELISA, sdf-1 Kit ELISA, sdf1 Kit ELISA, xSDF-1 Kit ELISA, SDF-1 Kit ELISA, cxcl12 Kit ELISA, sdf-1a Kit ELISA, sdf1a Kit ELISA, wu:fa55e10 Kit ELISA, wu:fc16h12 Kit ELISA, wu:fj84c02 Kit ELISA, C-X-C motif chemokine ligand 12 Kit ELISA, C-X-C motif chemokine ligand 1 Kit ELISA, chemokine (C-X-C motif) ligand 12 Kit ELISA, chemokine (C-X-C motif) ligand 1 Kit ELISA, C-X-C motif chemokine ligand 12 L homeolog Kit ELISA, chemokine (C-X-C motif) ligand 12a (stromal cell-derived factor 1) Kit ELISA, CXCL12 Kit ELISA, cxcl12 Kit ELISA, Cxcl1 Kit ELISA, Cxcl12 Kit ELISA, cxcl12.L Kit ELISA, cxcl12a Kit ELISA

Sujet: SDF-1 (Stromal cell-derived factor-1), also known as PBSF (pre-B-cell growth-stimulating factor), is a recently discovered protein belonging to the alpha chemokine (CXC) family of cytokines. SDF-1alphaand SDF-1beta are the first cytokines initially identified using the signal sequence trap cloning strategy from a human bone-marrow stromal cell line. SDF-1 has chemotactic activity on resting T lymphocytes and monocytes. The SDF-1alphaELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human SDF-1alphain plasma ( serum samples are not recommended for use in this assay) cell culture supernatants and urine. This assay employs an antibody specific for human SDF-1alphacoated on a 96-well plate. Standards and samples are pipetted into the wells and SDF-1alphapresent in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human SDF-1alphaantibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of SDF-1alphabound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.

ID gène: 6387

UniProt: P48061

Pathways: Regulation of Cell Size, CXCR4-mediated Signaling Events, Negative Regulation of intrinsic apoptotic Signaling