The Rat hypoxia-inducible factor 1α (HIF-1α) ELISA kit is a ready-to-use microwell, strip plate ELISA kit for the quantitative measurement of rat HIF-1α in biological samples, including serum, plasma, and tissue homogenates. It employs the sandwich ELISA technique and enzyme-substrate chromogenic reaction in the detection of target analytes in the samples. This ELISA kit has been quality controlled in multiple characteristics, including sensitivity, specificity, precision, linearity, and recovery. See the product instructions to obtain more information.
HIF-1α, a basic-helix-loop-helix (bHLH)-PER (Period)-ARNT-SIM (single-minded) (PAS) superfamily member, is the O2-labile HIF-1 subunit and is widely expressed in hypoxia. HIF-1α orchestrates cellular adaptation to low oxygen and nutrient-deprived environment and drives progression to malignancy in human solid cancers. Under normal O2 tension, the prolyl hydroxylases (PHDs) become active and hydroxylate HIF-1α, triggering von Hippel–Lindau (pVHL)-mediated ubiquitination and proteasomal degradation HIF-1. Hypoxia inhibits the enzymatic activity of PHDs, terminating proline modification and pVHL-HIF binding, causing HIF-1α stabilization, nuclear translocation, and further formation of HIF-1 complex. This cascade of events ultimately induces the transcription of genes that promote glycolytic metabolism, angiogenesis, and survival. HIF-1α is physiologically activated during embryogenesis and in wound-healing processes. Whereas in cancer, HIF-1α activation is associated with malignancy and poor prognosis.
||hypoxia inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor)
||Hif1aHypoxia-inducible factor 1-alpha ELISA Kit; HIF-1-alpha ELISA Kit; HIF1-alpha ELISA Kit
||Rattus norvegicus (Rat)
||serum, plasma, tissue homogenates
||3.12 pg/mL-200 pg/mL
||Epigenetics and Nuclear Signaling
|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. || || |
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|To assess the linearity of the assay, samples were spiked with high concentrations of rat HIF-1α 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)|| |
|1:1||Average %||90|| |
|Range %||87-94|| |
|1:2||Average %||96|| |
|Range %||91-100|| |
|1:4||Average %||93|| |
|Range %||87-97|| |
|1:8||Average %||100|| |
|Range %||96-103|| |
|The recovery of rat HIF-1α 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) ||92||87-95|| |
|EDTA plasma (n=4)||95||88-98|| |
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|These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed. |
|200||2.715 ||2.656 ||2.686 ||2.538 || |
|100||1.730 ||1.676 ||1.703 ||1.555 || |
|50||1.196 ||1.179 ||1.188 ||1.040 || |
|25||0.608 ||0.618 ||0.613 ||0.465 || |
|12.5||0.404 ||0.412 ||0.408 ||0.260 || |
|6.25||0.303 ||0.316 ||0.310 ||0.162 || |
|3.12||0.248 ||0.237 ||0.243 ||0.095 || |
|0||0.150 ||0.146 ||0.148 || || |
- A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-Rat HIF-1α 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 HIF-1α 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