脂滴螢光檢測試劑盒(Lipid Droplets Fluorescence Assay Kit) | SERVICEBIO 貨號G1905-100T
SERVICEBIO細胞螢光染色(Cell Fluorescence Staining)系列產品- BODIPY 493/503
脂滴螢光檢測試劑盒(Lipid Droplets Fluorescence Assay Kit) | SERVICEBIO 貨號G1905-100T
(4,4-Difluoro-1,3,5,7,8-Pentamethyl-4-Bora-3a,4a-Diaza-s-Indacene)
產品介紹:
脂滴(Lipid droplets)是細胞內特化的脂質儲存細胞器，可調節細胞內中性脂質的儲存(storage)和水解(hydrolysis)，為細胞提供動態的能量平衡。此外，脂滴(Lipid droplets)能夠沿著細胞骨架(cytoskeleton)移動並與其他細胞器相互作用，在訊號傳導(signal transduction)的調節中發揮重要作用。細胞脂滴( lipid droplets)的異常累積發生在多種代謝疾病中，如肥胖(obesity)、脂肪肝(fatty liver)、心血管疾病(cardiovascular diseases)和病理狀態(pathological states)。
脂滴螢光檢測試劑盒(Lipid Droplets Fluorescence Assay Kit, G1905-100T) 基於BODIPY 493/503，一種能夠定位極性脂質的親脂性螢光探針，可用於校準細胞和組織的中性脂質含量，該試劑盒經優化和調試，具有使用方便、性能穩定、重現性好等多重優勢。細胞內脂質含量越高，產生的螢光強度越強，可透過螢光顯微鏡(fluorescence microscope )和流式細胞儀(flow cytometer)檢測。此外，該試劑盒還額外提供了油酸( oleic acid)作為三酸甘油酯(triglyceride )合成和儲存的誘導劑，可用於陽性對照誘導(positive control induction)。
| Component Number | Component | G1904-100T |
| G1905-1 | Lipid droplet assay probe (2 mM) | 100 μL |
| G1905-2 | Lipid Droplet Assay Buffer | 100 mL |
| G1905-3 | Positive inducer (500 mM) | 50 μL |
| Manual | 1 pc | |
Assay Protocol
Note: Take 6-well plate adherent cells as an example, suspension cells need to be centrifuged every time the liquid is changed; Tissue samples directly refer to the staining and labeling steps, and the overall volume of reagents used is adjusted according to the situation; If necessary, the samples can be fixed in advance, but please avoid perforation of the samples.
1. Preparation of working solution for lipid droplet assay and working solution for lipid droplet induction:
1.1. Take appropriate amount of lipid droplet assay probe (2 mM) and lipid droplet assay buffer, mix thoroughly at a ratio of 1 : 500-2000, and prepare a working solution for lipid droplet assay;
1.2. Take an appropriate amount of positive inducer (500 mM) and use the cell culture medium of the corresponding test cells to dilute it to 400-1000 uM, and prepare a lipid droplet induction working solution for spare use (this concentration is a reference range, and can be adjusted appropriately according to the nature of the cells or other circumstances).
2. Positive control group and drug and other treatments (6-well plate adherent cells, for example, suspension cells each time the change of liquid, please perform centrifugation):
2.1. Cells are pre-planted in 6-well plates as needed (with or without crawler depending on assay and equipment);
2.2. Remove the original cell culture medium and wash 2 times with PBS or other buffers;
2.2.1. Experimental group: the cells to be tested are subjected to drug stimulation or other relevant treatments, and the desired incubation time is set by yourself;
2.2.2. Positive control group: 1 mL of lipid droplet induction working solution is added to the well plate, which is placed in an incubator for overnight incubation;
2.3. At the end of the incubation, turn to step 3 for stain labeling.
3. Lipid droplet staining marker:
3.1. Remove the cellogenic medium and wash 2-3 times with PBS or other buffers;
3.2. Remove the wash buffer, add 1 mL of Lipid Droplet Assay Working Solution and mix with gentle shaking;
3.3. Incubate for 15 min in a CO2 incubator protected from light;
3.4. After incubation, wash it twice with PBS or other buffers, and it can be further stained with nuclei and other operations or detected with the appropriate equipment according to the experimental needs;
4. Lipid droplet detection:
4.1. The sample after incubation and washed in 3.4 is analyzed using the appropriate method and instrumentation, labeled to show green fluorescence, EX/EM 493/503 nm, and some examples of instrumental detection are given below;
4.1.1. Directly or with anti-fluorescence quenching sealing solution (Item No. G1401), the film is sealed and then placed under a fluorescence microscope for detection and analysis.
4.1.2. After digestion and resuspension using tryptic digest (suspension cells are not used), cells are detected and analyzed using flow cytometry.
Fluorescence Microscopy to Observe The Procedure of Living Dead Cells：
1. Due to the different types and states of the cells, the working concentration of the dye and the incubation time can be adjusted appropriately.
2. The dye needs to be used for mitochondrial localization in the state of living cells, and the cells cannot be fixed first, otherwise it will lead to false positive results of inaccurate localization.Fixation and subsequent operations can be performed after staining of living cells.
3. When staining and washing live cells, the assay working solution and washing buffer need to be pre-warmed at 37°C. Staining and washing are accomplished by incubation in an incubator so as not to affect the morphology of the cells by temperature difference stimulation.
4. In order to prevent fluorescence quenching, the entire staining process needs to be operated away from light.
5. Probe master mixes should be stored in separate containers to avoid repeated freezing and thawing.To avoid waste, the staining solution is prepared as needed.
6. For your health and safety, please wear lab coat and gloves during operation.
流式細胞儀偵測細胞週期程序：
註：以6孔板貼壁細胞為例，懸浮細胞每次換液需離心；組織樣本直接參考染色、標記步驟，整體試劑用量依狀況調整；如有必要，可提前固定樣品，但請避免樣品穿孔。
1.脂滴測定工作液及脂滴誘導工作液的製備：
1.1.取脂滴檢測探針（2 mM）及脂滴檢測緩衝液適量，以1:500-2000的比例充分混合，製備脂滴檢測工作液；
1.2.取適量陽性誘導劑（500 mM），以相應測試細胞的細胞培養基稀釋至400-1000 uM，配製脂滴誘導工作液備用（此濃度為參考範圍，並可依細胞性質或其他情況適當調整） 。
2.陽性對照組及藥物等處理（以6孔板貼壁細胞為例，懸浮細胞每次換液時請進行離心）：
2.1.根據需要將細胞預先種植在 6 孔板中（根據檢測和設備，有或沒有爬行器）；
2.2.除去原始細胞培養基，以PBS或其他緩衝液洗滌2次；
2.2.1.實驗組：待測細胞接受藥物刺激或其他相關處理，所需孵育時間自行設定；
2.2.2.陽性對照組：在孔板中加入1mL脂滴誘導工作液，置於培養箱中孵育過夜；
2.3.孵育結束時，轉至步驟 3 進行染色標記。
3. 脂滴染色標記：
3.1.除去細胞生成培養基，以PBS或其他緩衝液洗滌2-3次；
3.2.除去洗滌緩衝液，加入1 mL 脂滴測定工作液，輕輕搖晃混勻；
3.3. CO2培養箱避光孵育15分鐘；
3.4.孵育後以PBS或其他緩衝液洗滌兩次，可依實驗需要進一步進行細胞核染色等操作或以適當的設備進行檢測；
4.脂滴檢測：
4.1. 3.4中孵育和洗滌後的樣品使用適當的方法和儀器進行分析，標記為顯示綠色螢光，EX/EM 493/503 nm，下面給出一些儀器檢測的例子；
4.1.1.直接或以抗螢光淬滅封閉液（貨號G1401）將膜封閉後置於螢光顯微鏡下檢測分析。
4.1.2.使用胰蛋白酶消化進行消化和重懸（不使用懸浮細胞）後，使用流式細胞儀檢測和分析細胞。
螢光顯微鏡觀察活死細胞的過程：
1.由於細胞類型和性質不同，陽性誘導劑和探針的濃度以及孵育時間可根據實驗需要適當調整。
2. 樣品應避免與有機溶劑或其他影響脂質的成分接觸，以免干擾測定的準確性。
3.螢光染料需淬滅，過程應避光。
4.為了您的健康與安全，操作時請穿戴實驗服並戴上手套。
|
Cat.No.
|
Product Name
|
Spec.
|
|---|
|
G1011-10ML
|
Hoechst 33258 Staining Solution (Ready-To-Use)
Hoechst 33258 染色液（即用型） |
10 mL
|
|
G1012-100ML
|
DAPI Staining Reagent (Ready to Use)
DAPI 染色試劑（即使用型） |
100 mL
|
|
G1012-10ML
|
DAPI Staining Reagent (Ready to Use)
DAPI 染色試劑（即使用型） |
10 mL
|
|
G1021-10ML
|
PI Stain
碘化丙啶(propidium iodide, PI) |
10 mL
|
|
G1127-1ML
|
Hoechst 33342 Staining Solution
Hoechst 33342 染色液 |
1 mL
|
|
G1515-100T
|
JC-1 Mitochondrial Membrane Potential Assay Kit
JC-1 粒線體膜電位檢測試劑盒 |
100 T
|
|
G1704
|
DiO Cell Membrane Green Fluorescence Staining Kit
DiO 細胞膜綠色螢光染色試劑盒 |
100-1000 T
|
|
G1705
|
Dil Cell Membrane Red Fluorescence Staining Kit
Dil 細胞膜紅色螢光染色試劑盒 |
100-1000 T
|
|
G1720-20UL
|
ER-Tracker Green
ER-追蹤器綠色(ER-Tracker Green) |
20 μL
|
|
G1721-20UL
|
ER-Tracker Red
ER-追蹤器紅色(ER-Tracker Red) |
20 μL
|
|
G1722-50UL
|
Lyso-Tracker Green
溶血追蹤器綠色(Lyso-Tracker Green) |
50 μL
|
|
G1723-50UG
|
MitoTracker Red CMXRos
MitoTracker 紅色 |
50 μg
|
|
G1905-100T
|
Lipid Droplets Fluorescence Assay Kit
脂滴螢光檢測試劑盒 |
100 T
|
Servicebio台灣品牌介紹-BRAND/AGENCY/品牌介紹- Servicebio
| Classfication | Cat No. | Name | Place of Origin | Spec. |
|
Amino Acids
|
GC304002 | L-Glutamine | 產品資料 | 100 g |
| GC304003 | L-Arginine hydrochloride | 產品資料 | 25 g | |
| GC304004-100g | L-Glutamine acid | 產品資料 | 100 g | |
| GC304005 | L-Isoleucine | 產品資料 | 25 g | |
| GC304006-25g | L-Leucine | 產品資料 | 25 g | |
| GC304007 | L-Lysine hydrochloride | 產品資料 | 25 g | |
| GC304009 | L-Methionine | 產品資料 | 25 g | |
| GC304010 | L-Phenylalanine | 產品資料 | 10 g | |
| GC304011-25g | L-Alanine | 產品資料 | 25 g | |
| GC304012 | L-Histidine | 產品資料 | 25 g | |
| GC304013 | L-Serine | 產品資料 | 25 g | |
| GC304014 | L-tryptophan | 產品資料 | 5 g | |
| GC304016 | L-Threonine | 產品資料 | 25 g | |
| GC304017 | L-Valine | 產品資料 | 10 g | |
| GC304019 | Glycine | 產品資料 | 500 g | |
| GC304021 | L-Histidine hydrochloride monohydrate | 產品資料 | 25 g | |
| GC304022 | Glycine cell culture grade 甘氨酸Glycine (細胞培養等級 |
產品資料 | 100 g | |
|
Protein Derivatives
|
GC305002-5g | Iodoacetamide | 產品資料 | 5 g |
| GC305003 | Trypsin 1:250 Source: Porcine Pancreas | 產品資料 | 10 g | |
| GC305004-5g | Pepsin 1:3000 | 產品資料 | 5 g | |
| GC305005 | Horseradish peroxidase | 產品資料 | 1 g | |
| GC305006 | Bovine Serum Albumin – import | 產品資料 | 100 g | |
| GC305009 | Human Albumin-FITC | 產品資料 | 0.5 mL | |
| GC305010 | Bovine Serum Albumin BSA | 產品資料 | 100 g | |
| GC305010-25G | Bovine Serum Albumin BSA | 產品資料 | 25 g | |
| GC305010-50G | Bovine Serum Albumin BSA | 產品資料 | 50 g | |
| GC305010-5G | Bovine Serum Albumin BSA | 產品資料 | 5 g | |
| GC305013-100mg | Collagenase I | 產品資料 | 100 mg | |
| GC305014-100mg | Collagenase II | 產品資料 | 100 mg | |
| GC305015-100mg | Collagenase IV 膠原酶IV (Collagenase IV) |
產品資料 | 100 mg | |
| GC305016-100g | Gelatine | 產品資料 | 100 g | |
|
Hormones
|
GC302002-25mg | Abscisic acid | 產品資料 | 25 mg |
| GC302003 | Angiotensin II | 產品資料 | 10 mg | |
|
Biological pigments
|
GC307002-25g | Crystal violet | 產品資料 | 25 g |
| GC307003-5g | Trypan Blue | 產品資料 | 5 g | |
| GC307005 | Alcian Blue8GX | 產品資料 | 1 g | |
| GC307006-5g | Bromphenol Blue | 產品資料 | 5 g | |
| GC307007 | Serva blue G-250 | 產品資料 | 10 g | |
| GC307008-5g | Congo Red | 產品資料 | 5 g | |
| GC307009-25g | Alizarin Red S | 產品資料 | 25 g | |
| GC307010-1g | Giemsa′s Stain | 產品資料 | 1 g | |
| GC307011 | Fuchsin basic | 產品資料 | 5 g | |
| GC307012-10g | Ponceau S | 產品資料 | 10 g | |
| GC307013 | Acid red 26 2R | 產品資料 | 5 g | |
| GC307014 | Direct Red 80 | 產品資料 | 5 g | |
| GC307015-5g | Sudan Black B | 產品資料 | 5 g | |
| GC307017 | Aniline blue | 產品資料 | 5 g | |
| GC307018 | Nuclear fast red | 產品資料 | 5 g | |
| GC307019 | Solvent Blue 38 | 產品資料 | 5 g | |
| GC307020 | Hematoxylin | 產品資料 | 5 g | |
| GC307021 | Solvent Red 43 | 產品資料 | 10 g | |
| GC307022 | Eosin Y, water soluble | 產品資料 | 10 g | |
| GC307023 | Safranin O | 產品資料 | 10 g | |
| GC307024 | Fast Green | 產品資料 | 10 g | |
| GC307026-5g | Pyronin G | 產品資料 | 5 g | |
| GC307027-1g | Janus Green B | 產品資料 | 1 g | |
| GC307030-10g | Brilliant Cresyl Blue | 產品資料 | 10 g | |
| GC307031 | Coomassie brilliant blue R-250 | 產品資料 | 5 g | |
| GC307034-1g | Thionine acetate | 產品資料 | 1 g | |
| GC307035-1g | Cresyl Fast Violet | 產品資料 | 1 g | |
| GC307036-1g | Orcein | 產品資料 | 1 g | |
| GC307037-1g | Carmine | 產品資料 | 1 g | |
| GC307039-10g | Mordant Blue 29 | 產品資料 | 10 g | |
| GC307042-5g | Oil Red O | 產品資料 | 5 g | |
| GC307045-25g | Rhodamine B | 產品資料 | 25 g | |
|
Vitamins
|
GC301015-25g | Vitamin C | 產品資料 | 25 g |
| GC303001 | Folic acid | 產品資料 | 5 g | |
| GC303004 | Ribofavin | 產品資料 | 25 g | |
| GC303005-5g | VitaminB6 | 產品資料 | 5 g | |
| GC303006-25g | VitaminB3 | 產品資料 | 25 g | |
| GC303008 | VitaminB12 | 產品資料 | 1 g | |
| GC303011-25mg | NADPH-Na4 | 產品資料 | 25 mg | |
| GC303013-250mg | β-nicotinamide adenine dinucleotide NAD | 產品資料 | 250 mg | |
| GC303014-250mg | NADH-Na2 | 產品資料 | 250 mg | |
|
Antibiotics
|
GC301002 | Streptozocin STZ | 產品資料 | 1 g |
| GC301002-100mg | Streptozocin STZ | 產品資料 | 100 mg | |
| GC301003 | Penicillin G Sodium salt | 產品資料 | 1 g | |
| GC301004 | Ampicillin | 產品資料 | 1 g | |
| GC301005 | Streptomycin sulfate | 產品資料 | 5 g | |
| GC301008 | Kanamycin | 產品資料 | 1 g | |
| GC301009 | Kanamycin sulfate | 產品資料 | 5 g | |
| GC301009-1g | Hygromycin B | 產品資料 | 1 g | |
| GC301011 | Geneticin G-418 | 產品資料 | 1 g | |
| GC301013 | Chloramphenicol | 產品資料 | 5 g | |
| GC301013-10mg | Pseudomonic acid | 產品資料 | 10 mg | |
| GC301014-5g | Norfloxacin | 產品資料 | 5 g | |
| GC301016 | Cisplatin | 產品資料 | 100 mg | |
| GC301017-100mg | Sirolimus | 產品資料 | 100 mg | |
|
Heterocyclic compounds
|
GC207001 | Imidazole | 產品資料 | 10 g |
| GC207002 | PMSF | 產品資料 | 5 g | |
| GC207003 | Freund’s Adjuvant Complete | 產品資料 | 10 mL | |
| GC207004 | Freund’s Adjuvant Incomplete | 產品資料 | 10 mL | |
|
Salt
|
GC201001 | Guanidine HCl | 產品資料 | 500 g |
| GC201002 | Guanidine thiocyanate | 產品資料 | 500 g | |
|
Sugar Alcohols
|
GC205001 | L(+)-Lactic acid | 產品資料 | 100 g |
| GC205002 | Pyruvic acid sodium salt | 產品資料 | 25 g | |
| GC205003 | D-Glucose, anhydrous | 產品資料 | 500 g | |
| GC205004-10g | D(+)-Trehalose dihydrate | 產品資料 | 10 g | |
| GC205005 | D(+)- Galactose | 產品資料 | 100 g | |
| GC205006-10g | D-(-)Arabinose | 產品資料 | 10 g | |
| GC205007-1g | DTE | 產品資料 | 1 g | |
| GC205008 | Glycerol | 產品資料 | 100 mL | |
| GC205009 | LPS | 產品資料 | 10 mg | |
| GC205010 | DTT | 產品資料 | 1 g | |
| GC205011 | IPTG | 產品資料 | 1 g | |
| GC205013 | Agarose | 產品資料 | 100 g | |
| GC205014 | Sucrose | 產品資料 | 500 g | |
| GC205015 | D-Sorbitol | 產品資料 | 500 g | |
|
Solvent
|
GC203001 | TEMED | 產品資料 | 100 mL |
| GC203002 | DMSO(Dimethyl sulfoxide) | 產品資料 | 100 mL | |
| GC203003 | Urea | 產品資料 | 500 g | |
| GC203005 | DMSO cellular level | 產品資料 | 10 mL | |
|
Complex
|
GC202001 | EDTA | 產品資料 | 500 g |
| GC202002 | EDTA -Na2 | 產品資料 | 500 g | |
|
Buffer salt
|
GC208001 | Tris (Hydroxymethyl) Aminomethane | 產品資料 | 500 g |
| GC208003 | RIS-HCL | 產品資料 | 500 g | |
| GC208004-100g | BIS-TRIS | 產品資料 | 100 g | |
| GC208006 | N-Tris[Hydroxymethyl]Methylglycine | 產品資料 | 100 g | |
| GC208007 | 4-propanesulfonyl morpholine | 產品資料 | 100 g | |
| GC208008-25g | HEPPS | 產品資料 | 25 g | |
| GC208009 | HEPES, Free acid | 產品資料 | 100 g | |
| GC208012-1g | TCEP·HCL | 產品資料 | 1 g | |
|
Surfactant
|
GC204001 | Tween-80 | 產品資料 | 100 mL |
| GC204002 | Tween-20 | 產品資料 | 100 mL | |
| GC204003 | Triton X-100 | 產品資料 | 100 mL | |
| GC204004 | Sodium dodecylsulfate | 產品資料 | 500 g | |
| GC204005 | Sodium dodecylsulfate | 產品資料 | 100 g | |
| GC204006 | Nonidet TM P40 Substitute NP-40 | 產品資料 | 100 g | |
| GC204007 | Deoxycholic acid， sodium salt | 產品資料 | 25 g | |
| GC204009-100ml | APES | 產品資料 | 100 mL | |
|
Amine organics
|
GC206002 | Acrylamide | 產品資料 | 500 g |
| GC206003 | N’-MBA | 產品資料 | 100 g | |
|
Polymer
|
GC209002-250g | PEG 8000 | 產品資料 | 250 g |
| GC209003-250g | PEG 6000 | 產品資料 | 250 g | |
| GC209004-100g | PEG 3350 | 產品資料 | 100 g | |
| GC209005-500g | Poly(ethylene glycol)2000 | 產品資料 | 500 g | |
| GC209006-250g | Poly(ethylene glycol) 20000 | 產品資料 | 250 g | |
|
Inorganic salt
|
GC106001 | Orthoboric acid | 產品資料 | 500 g |
| GC104004 | Sodium bicarbonate | 產品資料 | 500 g | |
| GC108003 | Citric acid monohydrate | 產品資料 | 100 g | |
| GC108004 | Sodium citrate dihydrate | 產品資料 | 100 g | |
| GC102001 | Calcium chloride | 產品資料 | 100 g | |
| GC102002 | Magnesium chloride hexahydrate | 產品資料 | 100 g | |
| GC102003 | Potassium chloride | 產品資料 | 500 g | |
| GC102004 | Sodium chloride | 產品資料 | 500 g | |
| GC101004 | Magnesium Sulfate | 產品資料 | 100 g | |
| GC101013 | Ammonium persulfate | 產品資料 | 25 g | |
| GC105001 | Sodium phosphate monobasic | 產品資料 | 500 g | |
| GC105002 | Sodium Phosphate, Dibasic | 產品資料 | 500 g | |
| GC105003 | Potassium Phosphate Monobasic | 產品資料 | 500 g | |
| GC107001 | Sodium hydroxide | 產品資料 | 100 g | |
| GC306001 | Lodixanol | 產品資料 | 5 g | |
| GC310001-100g | skimmed milk powder | 產品資料 | 100 g |