Introduction
Immuno Fluorence Staining Kits are developed for immunofluorescence detection of cell or tissue sections. When there is an appropriate antibody to detect specific target protein, fluorescence can be detected by immunofluorescence staining kit.
Immuno Fluorescence Staining Kit (Anti-Rabbit IgG-Cyanine3) contains Goat Anti-Rabbit IgG(H+L)(Cyanine3 conjugated), this secondary antibody can detect primary antibody from rabbit with bright red fluorescence.
Cyanine3 is a newly red fluorescent probe. It is brighter than most of other red fluorescent probes, more difficult to quench, and has a lower background.
The kit contains anti fluorescence quenching sealing solution, which can make the fluorescence more lasting.
Components
|
Cat.
|
Product
|
50
Assays
|
100
Assays
|
200
Assays
|
Storage
|
|
E-AB-1010
|
Goat Anti-Rabbit IgG(H+L)( Cyanine3 conjugated)
|
120 μL
|
200 μL
|
200 μL×2
|
-20°C
|
|
E-IR-R110
|
Normal Goat
Blocking Buffer (Ready-to-Use)
|
5 mL
|
10 mL
|
20 mL
|
2~8°C
|
|
E-IR-R103
|
DAPI Reagent (1 μg/mL)
|
5 mL
|
10 mL
|
20 mL
|
2~8°C
|
|
E-IR-R119
|
Anti-Fluorescence
Quenching Agent
|
5 mL
|
10 mL
|
20 mL
|
2~8°C
|
|
Manual
|
One
copy
|
Preparation of Immunofluorescence Staining
A. Preparation of Fixation Solution
It is recommended to use 4% Paraformaldehyde as the fixation solution (E-IR-R113), or use ethanol, methanol or other types of fixative according to specific primary antibody or sample.
B. Preparation of Permeate working solution
Use Triton X-100 as the permeable solution
(E-IR-R122) and dilute with 1x PBS buffer to 0.5% Triton X-100 working
solution.
C. Preparation of TBST Working Buffer
It is recommended to use TBST as washing buffer. Use Elabscience ® 10× TBST (E-BC-R335) and dilute to 1 ×TBST Working Buffer with deionized water at ratio of 1:9.
D. Preparation of Antibody Dilution Solution
It is recommended to use Elabscience ® Antibody Dilution Buffer (E-IR-R106) or PBS as primary antibody dilution buffer.
E. Dilute Primary Antibody
Dilute the primary antibody according to the manual of primary antibody.
F. Dilute the Secondary Antibody
It is recommended to use Elabscience ® Antibody Dilution Buffer (E-IR-R106) or PBS as secondary antibody dilution buffer. Dilute the secondary antibody with antibody dilution buffer at the dilution of 1:50. The dilution ratio can be increased or decreased appropriately according to the intensity of fluorescence.
Storage
Store at 2~8/-20°C, Refer to the label. Valid for 12 months. Secondary antibody (E-AB-1010) and DAPI Reagent (E-IR-R103) should be stored at -20°C and avoid light.
Notice
1. This result should be observed by fluorescence microscope.
2. Anti-Fluorescence Quenching Reagent can slow down the quenching, but avoid light and especially shorten the result observation time is still needed.
3. If it can't be observed in time, please store the slice at 4°C and avoid light and observe in one week.
4. If the fluorescence is too weak, increase the primary antibody concentration properly. If the fluorescence is still weak, increase the secondary antibody concentration appropriately.
5. The reagents for immunofluorescence staining and the cover glass and slides should be prepared in advance.
6. For your safety and health, please wear the lab coat and disposable gloves before the experiments.
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