Biocytin |
Catalog No.GC14990
One-Click Copy Product Info
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Biocytin is a compound formed by the linkage of D-biotin and L-lysine via a secondary amide bond. Biocytin serves as a classic neuroanatomical tracer, utilizing its high-affinity binding to avidin for precise localization of neuronal morphology and connectivity.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 576-19-2
Sample solution is provided at 25 µL, 10mM.
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Biocytin is a compound formed by the linkage of D-biotin and L-lysine via a secondary amide bond. Biocytin serves as a classic neuroanatomical tracer, utilizing its high-affinity binding to avidin for precise localization of neuronal morphology and connectivity. Biocytin is primarily used in neural circuit mapping, biotinidase activity assays, and related diagnostic research[1-2].
References:
[1] McDonald AJ. Neuroanatomical labeling with biocytin: a review. Neuroreport. 1992 Oct;3(10):821-7.
[2] Ebrahim H, Dakshinamurti K. Determination of biocytin. Anal Biochem. 1987 May 1;162(2):319-24.
Biocytin Whole-Cell Recording and Tracing Method
This protocol is adapted from research data and provided for reference only. Adjustments may be necessary based on specific experimental requirements.
(1) Reagent and Electrode Preparation: Prepare intracellular solution containing 0.1% biocytin (components: 110mM K-gluconate, 10mM NaCl, 1mM MgCl₂, 10mM EGTA, 40mM HEPES, 2mM Mg-ATP, 0.3mM Na-GTP, pH=7.3, osmolarity 280-300mOsm). Control glass electrode resistance according to target neuron size (medium spiny neurons: 6-8MΩ; pyramidal neurons/parvalbumin interneurons: 5-6MΩ), ensuring the electrode tip is free of bubbles.
(2) Brain Slice Preparation and Neuron Localization: Obtain brain tissue from 3-month-old mice and prepare coronal slices containing the primary motor cortex (M1) or striatum (320μm thick) using a vibratome. After recovering in oxygenated artificial cerebrospinal fluid (aCSF) for 1 hour, transfer slices to a thermostatically controlled perfusion recording chamber (30°C). Identify neuronal morphology using infrared differential interference contrast (IR-DIC) microscopy (pyramidal neurons are teardrop-shaped, medium spiny neurons are oval-shaped).
(3) Whole-Cell Recording and Biocytin Loading: After the electrode contacts the cell membrane, apply negative pressure to form a gigaohm seal, then briefly apply strong suction to rupture the membrane and establish whole-cell mode. Record action potentials (AP) in current-clamp mode, then maintain whole-cell mode for 40-60 minutes to allow biocytin to fully diffuse into fine neuronal structures such as dendrites and axons. Slowly withdraw the electrode to allow the membrane to reseal.
(4) Tissue Fixation and Fluorescence Staining: Fix the recorded brain slices in 4% paraformaldehyde at 4°C for 24 hours. After permeabilization with 1% Triton-X100, incubate with streptavidin-Alexa 594 (red) or Alexa 488 (green) fluorescent probes (diluted 1:1000) at 4°C for 24 hours.
(5) Morphological Imaging and Analysis: Use a confocal microscope (63×oil immersion objective) to acquire Z-stack images of neurons (step size 0.2μm) under corresponding fluorescence signals. Perform 3D reconstruction using ImageJ software to analyze total dendritic length, branching complexity (Sholl analysis), and dendritic spine density (number of spines per unit length of dendrite).
Precautions:
(1) Experiments must comply with animal ethics guidelines.
(2) For your safety and health, please wear a lab coat and disposable gloves during operation.
References:
[1] Tan S, Mo X, Qin H, et al. Biocytin-Labeling in Whole-Cell Recording: Electrophysiological and Morphological Properties of Pyramidal Neurons in CYLD-Deficient Mice. Molecules. 2023 May 15;28(10):4092.
| Cas No. | 576-19-2 | SDF | |
| Chemical Name | (S)-2-amino-6-(5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)hexanoic acid | ||
| Canonical SMILES | O=C1N[C@H]([C@H](CCCCC(NCCCC[C@@H](C(O)=O)N)=O)SC2)[C@H]2N1 | ||
| Formula | C16H28N4O4S | M.Wt | 372.48 |
| Solubility | Water:35mg/ml | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.6847 mL | 13.4235 mL | 26.8471 mL |
| 5 mM | 536.9 μL | 2.6847 mL | 5.3694 mL |
| 10 mM | 268.5 μL | 1.3424 mL | 2.6847 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.50% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 18 reference(s) in Google Scholar.)