辅助产品-细胞膜染色试剂盒 Cell Plasma Membrane Staining Kit (Green Fluorescence)

Datasheet

Background & Principle

The cell membrane (plasma membrane) is a thin semi-permeable membrane, consisting of a lipid bilayer withembedded proteins that separates the interior of all cells from the environment. The basic function of the cellmembrane is to protect the cell from its surroundings. The cell membrane controls the movement of substances inand out of cells and organelles. In this way, it is selectively permeable to ions and organic molecules. Cellmembranes are involved in a variety of cellular processes such as cell adhesion, ion conductivity and cell signalingand serve as the attachment surface for several extracellular structures, including the cell wall, glycocalyx, andintracellular cytoskeleton. #A10020 Cell Plasma Membrane Staining Kits are a set of fluorescence imaging tools forrapid staining of plasma membranes in living and fixed suspended or attached cells depending on the cell type andexperimental conditions. The kit uses a proprietary lipophilic carbocyanine dye (Ex/Em = 484/501 nm) that weaklyfluorescent in water but highly fluorescent and quite photostable when incorporated into membranes.

Storage Stability

Refer to list of materials supplied for storage conditions of individual components. Stable for at least 6 months atrecommended temperature from date of shipment. Gel pack with blue ice.


Assay Restrictions

Assay kit is intended for research use only. Not for use in diagnostic procedures.

Do not mix or substitute reagents or materials from other kit lots or vendors. Kits are QC tested as a set ofcomponents and performance cannot be guaranteed if utilized separately or substituted.


Materials supplied and Storage conditions

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Other supplies required Not Supplied

  1. Microcentrifuge;

  2. 2. Pipettes and pipette tips;

  3. 3. Fluorescence Microscopy or Flow Cytometer;

4. 24-well plate for cell culture; 5. Phosphate-buffered saline (PBS);

Technical hints

ASSAY PROTOCOL

Reagent Preparation

PM Green (1000×): Warm to room temperature. Aliquot and store unused PM Green (1000×)stock solutions at -20°C. Protect from light and avoid repeated freeze-thaw cycles.Assay Buffer: Prepare 1×Assay Buffer by dilute 10× Assay Buffer with ddH2O. Warm to 37°Cbefore use.

Staining Solution: Mix 1 μl of PM Green (1000×) in each 1 ml of Assay Buffer. Scale up

accordingly for larger numbers of assays.Recommended procedures

Note: The optimal concentration of the PM Green and incubation time varies depending onthe specific application. The staining conditions may need modified according to the

particular cell type.

A Quantification by Flow Cytometry

1. Treat cells with the desired method.

Note: We recommend keeping unstained control cells (i.e. without PM Green) suspended inAssay Buffer for both treated and untreated samples to set up the flow cytometer instrument.

2. For non-adherent cells, Collect 1-5 ×105 cells by centrifugation (4oC, 300g, 5min). Washwith ice-cold PBS twice and discard the PBS. For adherent cells, using Trypsin (EDTA free) todigest cells firstly and then centrifugation.

3. Resuspend the cells pellet in 500uL Staining Solution.

4. Incubate the cells at 37°C for 5-20 minutes in the dark.

5. Centrifuge cells at 500g and discard supernatant.

6. Wash cell pellet with PBS and repeat step 5.

7. Resuspend cell pellet in 0.5 ml of the pre-warmed PBS and analyze the cells by flow

cytometry using FITC channel (usually FL1)

B Detection by Fluorescence Microscopy

1. For suspension cells: Follow the protocol for flow cytometry from step1 to step4 and placethe cell suspension from Step A.4 on a glass slide. Cover the cells with a glass coverslip.Analyze cells by fluorescence microscopy using the appropriate filters as soon as possible.

2. For adherent cells: the suggested protocol is as below.

2.1. Grow cells directly on a coverslip in 24 well dish. Incubate in a CO2 Incubator at 37°C forat least 24 hours before treatment.

2.2. Wash cells with PBS twice.

2.3. Add 0.5 mL of Staining solution to cells and incubate at 37°C for 5-10 minutes in the dark.

2.4. Wash cells with pre-warm PBS twice.

2.5. Fix cells after staining (Optional): Fix the cells with 4% paraformaldehyde for 15-30

minutes. Other fixatives, particularly glutaraldehyde, tend to produce unacceptably high levelsof background fluorescence.

2.6. Invert coverslip on a glass slide and visualize cells fluorescence microscopy using theappropriate filters as soon as possible (Ex/Em = 484/501 nm)

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A10020

货号 规格 价格(¥)
A10020T 100T

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Note:  Application as IHC, only suitable for histochemical staining or fluorescence staining of paraffin-embedded sections. Application as ICC/IF, suitable for histochemical or fluorescent staining of frozen sections, as well as chemical and fluorescent staining at the cellular level.

注意:抗体应用为IHC的,抗体只适合于石蜡切片的组化染色或者荧光染色。

抗体应用为IF/ICC的,抗体适合于冰冻切片的组化染色或者荧光染色,以及细胞水平的化学染色和荧光染色。

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微信客服

邮箱:market@ab-mart.com

应聘职位:hr@ab-mart.com

订购专线:4006-123-828

销售电话:13162017139(微信同号)

技术支持:13162477137(微信同号)

总机:021-34695901

经销商:QQ 402772198
南方经销商负责:手机13122837132(微信同号)          
北方及西南经销商负责:手机13122150513(微信同号)

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