Introduction, advantages, and operation steps of AF488 labeled ghost pen cyclic peptide
Ghost pen cyclic peptide is a cyclic peptide extracted from a fungus Amanita Dhalloides. It competes with F-actin binding. Fluorescent labeled ghost pen peptides can selectively bind to F-actin at nanogram levels and are easily soluble in water, providing a very convenient labeling method for detecting actin localization and quantification in tissue sections, cultured cells, and cell-free systems. The ghost pen cyclic peptide labeled with Alexa Fluor 488 can emit high brightness and stable green fluorescence, with strong staining reaction specificity and high contrast, making it suitable for qualitative and quantitative detection of F-actin.
Compared with Actin antibodies, Alexa Fluor 488 ghost pen peptide has the following advantages:
1. It binds one-to-one to actin subunits and does not bind to G-actin, resulting in better staining performance than Actin antibodies.
2. And the combination of this product has no species differences and has a wide range of applicability.
3. The non-specific signal of ghost pen cyclic peptide labeling can be ignored, resulting in good contrast in the image.
4. Staining is fully compatible with other fluorescent staining methods used for cell analysis, including fluorescent proteins, nanocrystals, and other Alexa Fluor conjugates (including Alexa Fluor conjugated secondary antibodies). After binding with this product, F-actin can still maintain many biological characteristics of actin itself
488 Ghost Pen Cyclic Peptide Staining Product Recommendation
[Brand] and Yuanliji Biotechnology
[Product] Tissue Cryopreservation Resuscitation Reagent
[Product Code] AC05L463
[Specification] 1 T
488 Ghost Pen Cyclic Peptide Staining Product Usage Method
Step 1: Prepare 1 x Alexa Fluor 488 labeled ghost pen peptide working solution. Transfer 1 μ L of 1000 x Alexa Fluor 488 labeled ghost pen peptide storage solution to 1 mL of PBS buffer containing 1% BSA to obtain 1 x working solution.
Step 2: Cell staining procedure
(1) Cell culture overnight or longer to achieve a density of 50-60% confluence.
(2) Remove the culture medium and wash the cells twice with 1 x PBS (pH 7.4) preheated at 37 ℃.
(3) Use PBS solution containing 3-4% formaldehyde for cell fixation, and fix at room temperature for 10-30 minutes.
(4) Wash the fixed cells with PBS 2-3 times at room temperature for 10 minutes each time.
(5) (Optional): At room temperature, permeabilize with 0.1% Triton X-100 solution dissolved in PBS for 3-5 minutes to increase its permeability.
(6) Wash cells 2-3 times with PBS at room temperature for 10 minutes each time.
(7) Add 100 μ L/well (96 well plate) of Alexa Fluor 488 labeled ghost pen cyclic peptide working solution to each well, and stain at room temperature in the dark for 20-90 minutes, with relatively shorter time in summer.
(8) Wash the cells three times with PBS, each time for 5 minutes, to remove excess Alexa Fluor 488 labeled ghost pen peptides.
(9) (Optional): Add sufficient ready to use DAPI solution for counterstaining of cell nuclei, such as 100 μ L/well (96 well plate), at room temperature for 3-5 minutes. Wash the cells twice with PBS for 5 minutes each time.
(10) Perform fluorescence observation under a fluorescence microscope or confocal microscope, using Alexa FluorTM 488 excitation/emission filter and/or DAPI excitation/emission filter (Ex/Im=364/454nm).
AF488 labeled ghost pen cyclic peptide can be used for labeling and detecting neuronal surface receptors. It can also be used to label and detect immune molecules on the surface of cells, thereby studying the activation and regulatory mechanisms of immune cells.
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