The Human stromal cell-derived factor 1 alpha (SDF1A) ELISA kit allows for the in vitro quantitative determination of SDF1A concentrations in multiple biological fluids, including serum, plasma, cell culture supernates, or tissue homogenates. It is not intended for diagnostic use. This assay kit was designed and optimized for immunology research use in humans. The kit has undergone rigorous quality control in multiple parameters, including sensitivity, specificity, precision, linearity, recovery, and inter-batch difference. Refer to the product instructions for more details.
This assay employs the quantitative sandwich enzyme immunoassay technique, in which SDF1A in the samples or standards are sandwiched between pre-coated SDF1A antibody and Biotin-conjugated SDF1A antibody. HRP-avidin is then added to the wells. Following a wash to remove any unbound reagent, the TMB substrate solution is added to the wells and color develops in proportion to the amount of SDF1A bound in the initial step. The color development is stopped upon adding the stop solution, and the intensity of the color is measured at 450 nm via a microplate reader. The levels of SDF1A in the samples can be determined by referring to the O.D. (optical density) of the samples to the standard curve.
SDF1A is the most abundant SDF1 isoform in the brain and is expressed by stromal cells. It is retained in the marrow by heparans and proteoglycans. The SDF1A/CXCR4 interaction is a migrational stimulus and leads to the retention of progenitors. Interfering with interactions of SDF1A and CXCR4 blocks the in vivo homing of hemopoietic stem cells (HPSCs) to the bone marrow (BM) and mobilizes stem cells for release into the blood. SDF1A ligation has also been involved in the arrest of progenitor cell cycling, which facilitates a transition into quiescence.
||stromal cell-derived factor 1 alpha (SDF1A)
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||Homo sapiens (Human)
||serum, plasma, cell culture supernates, tissue homogenates
||31.25 pg/mL-2000 pg/mL
|Intra-assay Precision (Precision within an assay): CV%<8%|| || || |
|Three samples of known concentration were tested twenty times on one plate to assess. || |
|Inter-assay Precision (Precision between assays): CV%<10%|| || || |
|Three samples of known concentration were tested in twenty assays to assess. || || |
| || || || || || || |
|To assess the linearity of the assay, samples were spiked with high concentrations of human SDF1A in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.|
| ||Sample||Serum(n=4)|| |
|1:5||Average %||88|| |
|Range %||85-92|| |
|1:10||Average %||96|| |
|Range %||91-102|| |
|1:20||Average %||92|| |
|Range %||86-98|| |
|1:40||Average %||90|| |
|Range %||85-96|| |
|The recovery of human SDF1A spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.|
|Sample Type||Average % Recovery||Range|| |
|Serum (n=5) ||93||87-96|| |
|EDTA plasma (n=4)||95||90-99|| |
| || || || || || || |
| || || || || || || |
|These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed. |
|2000||2.750 ||2.732 ||2.741 ||2.640 || |
|1000||2.144 ||2.136 ||2.140 ||2.039 || |
|500||1.163 ||1.144 ||1.154 ||1.053 || |
|250||0.630 ||0.628 ||0.629 ||0.528 || |
|125||0.348 ||0.324 ||0.336 ||0.235 || |
|62.5||0.260 ||0.246 ||0.253 ||0.152 || |
|31.25||0.210 ||0.201 ||0.206 ||0.105 || |
|0||0.105 ||0.097 ||0.101 || || |
- A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human SDF1A antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
- Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
- One vial Biotin-labeled SDF1A antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
- One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
- One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
- One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
- One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
- One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
- One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
- One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
- Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
- An instruction manual
|Materials not provided
- A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
- An incubator can provide stable incubation conditions up to 37°C±5°C.
- Squirt bottle, manifold dispenser, or automated microplate washer
- Absorbent paper for blotting the microtiter plate
- 50-300ul multi-channel micropipette
- Pipette tips
- Single-channel micropipette with different ranges
- 100ml and 500ml graduated cylinders
- Deionized or distilled water
- Test tubes for dilution
|ELISA kit FAQs
||Store at 2-8°C. Please refer to protocol.
||3-5 working days