- ￥2600
- life
- 11668-027
- 2025年08月30日
12 年金牌会员
企业认证
相关产品推荐更多 >
万千商家帮你免费找货
0 人在求购买到急需产品
- 详细信息
- 询价记录
- 文献和实验
- 技术资料
- 英文名：
Lipofectamine 2000
- 库存：
2
- 规格：
0.75ml
Lipofectamine™ 2000
Cat. No. 11668-027 Size: 0.75 ml Cat. No. 11668-019 Size: 1.5 ml Cat. No. 11668-500 Size: 15 ml
Store at +4°C (do not freeze)
Description
Lipofectamine™ 2000 is a proprietary formulation for the transfection of nucleic acids (DNA and RNA) into eukaryotic cells providing the following advantages:
? Highest transfection efficiency in many cell types and formats (e.g. 96-well). Refer to the Cell Lines database at www.invitrogen.com for a list of cell types successfully transfected.
? Nucleic acid-Lipofectamine™ 2000 complexes can be added directly to cells in culture medium, in the presence or absence of serum.
? It is not necessary to remove complexes or change/add medium after transfection, but complexes may be removed after 4-6 hours.
Important Guidelines for Transfection
? Use the procedure on page 2 to transfect cells with short interfering RNA
(siRNA) or Stealth™ RNAi.
? Use the procedure on page 3 to transfect cells with plasmid DNA.
? We recommend Opti-MEM® I Reduced Serum Medium (Cat. No. 31985-062)
to dilute Lipofectamine™ 2000 and nucleic acids before complexing.
? Do not add antibiotics to media during transfection as this causes cell death.
? Maintain the same seeding conditions between experiments.
? Test serum-free media for compatibility with Lipofectamine™ 2000 since some serum-free formulations (e.g. CD293, SFM II, VP-SFM) may inhibit cationic lipid-mediated transfection.
Note: For more tips for your RNAi experiment, refer to “Seven Steps to RNAi Success”. This manual is available from www.invitrogen.com/rnai or Technical Service, as are
cell-type specific RNAi transfection protocols (see “RNAi protocols”.)
Quality Control
Lipofectamine™ 2000 is tested for absence of microbial contamination with blood agar plates, Sabaraud dextrose agar plates, and fluid thioglycolate medium, and functionally by transfection of CHO-K1 cells with a reporter plasmid.
Part No.: 11668.2k.pps Rev. Date: 11 July 2006
For research use only. Not intended for any animal or human therapeutic or diagnostic use.
For technical support, contact tech_service@invitrogen.com.
Stealth™ RNAi or siRNA Transfection
Page 2
Use this brief procedure to transfect Stealth™ RNAi or siRNA into mammalian cells in a 24-well format. For other formats, see Scaling Up or Down Transfections (page 4). All amounts and volumes are given on a per well basis. Use this procedure as a starting point; optimize transfections as described in Optimizing Stealth™ RNAi or siRNA Transfection, especially if you are transfecting a mammalian cell line for the first time.
1. One day before transfection, plate cells in 500 µl of growth medium without antibiotics such that they will be 30-50% confluent at the time of transfection. Note: Transfecting cells at a lower density allows a longer interval between transfection and assay time, and minimizes the loss of cell viability due to cell overgrowth.
2. For each transfection sample, prepare oligomer-Lipofectamine™ 2000
complexes as follows:
a. Dilute 20 pmol Stealth™ RNAi or siRNA oligomer in 50 µl Opti-MEM® I Reduced Serum Medium without serum (final concentration of RNA when added to the cells is 33 nM). Mix gently.
b. Mix Lipofectamine™ 2000 gently before use, then dilute 1 µl in 50 µl Opti- MEM® I Reduced Serum Medium. Mix gently and incubate for 5 minutes at room temperature. Note: Proceed to Step c within 25 minutes.
c. After the 5-minute incubation, combine the diluted oligomer with the diluted Lipofectamine™ 2000. Mix gently and incubate for 20 minutes at room temperature (solution may appear cloudy).
3. Add the oligomer-Lipofectamine™ 2000 complexes to each well containing cells and medium. Mix gently by rocking the plate back and forth.
Incubate the cells at 37°C in a CO2 incubator for 24-96 hours until you are ready to assay for gene knockdown. Medium may be changed after 4-6 hours.
Optimizing Stealth™ RNAi or siRNA Transfection
To obtain the highest transfection efficiency and low non-specific effects, optimize transfection conditions by varying RNA and Lipofectamine™ 2000
concentrations. Test 10-50 pmol RNA and 0.5-1.5 µl Lipofectamine™ 2000 for 24- well format. Depending on the nature of the target gene, transfecting cells at higher densities may also be considered when optimizing conditions.
Plasmid DNA Transfection
Page 3
Use the following procedure to transfect DNA into mammalian cells in a 24-well format. For other formats, see Scaling Up or Down Transfections (page 4). All amounts and volumes are given on a per well basis. Prepare complexes using a
DNA (µg) to Lipofectamine™ 2000 (µl) ratio of 1:2 to 1:3 for most cell lines. Transfect cells at high cell density for high efficiency, high expression levels, and to minimize cytotoxicity. Optimization may be necessary (see Optimizing Plasmid DNA Transfection, page 4).
1. Adherent cells: One day before transfection, plate 0.5-2 x 105 cells in 500 µl of growth medium without antibiotics so that cells will be 90-95% confluent at the time of transfection.
Suspension cells: Just prior to preparing complexes, plate 4-8 x 105 cells in
500 µl of growth medium without antibiotics.
2. For each transfection sample, prepare complexes as follows:
a. Dilute DNA in 50 µl of Opti-MEM® I Reduced Serum Medium without serum (or other medium without serum). Mix gently.
b. Mix Lipofectamine™ 2000 gently before use, then dilute the appropriate amount in 50 µl of Opti-MEM® I Medium. Incubate for 5 minutes at room temperature. Note: Proceed to Step c within 25 minutes.
c. After the 5 minute incubation, combine the diluted DNA with diluted
Lipofectamine™ 2000 (total volume = 100 µl). Mix gently and incubate for
20 minutes at room temperature (solution may appear cloudy). Note:
Complexes are stable for 6 hours at room temperature.
3. Add the 100 µl of complexes to each well containing cells and medium. Mix gently by rocking the plate back and forth.
4. Incubate cells at 37°C in a CO2 incubator for 18-48 hours prior to testing for transgene expression. Medium may be changed after 4-6 hours.
5. For stable cell lines: Passage cells at a 1:10 (or higher dilution) into fresh growth medium 24 hours after transfection. Add selective medium (if desired) the following day.
Optimizing Plasmid DNA Transfection
To obtain the highest transfection efficiency and low cytotoxicity, optimize transfection conditions by varying cell density as well as DNA and
Page 4
Lipofectamine™ 2000 concentrations. Make sure that cells are greater than 90%
confluent and vary DNA (µg): Lipofectamine™ 2000 (µl) ratios from 1:0.5 to 1:5.
Scaling Up or Down Transfections
To transfect cells in different tissue culture formats, vary the amounts of Lipofectamine™ 2000, nucleic acid, cells, and medium used in proportion to the relative surface area, as shown in the table. With automated, high-throughput systems, a complexing volume of 50 µl is recommended for transfections in 96- well plates. Note: You may perform rapid 96-well plate transfections by plating cells directly into the transfection mix. Prepare complexes in the plate and directly add cells at twice the cell density as in the basic protocol in a 100 µl volume. Cells will adhere as usual in the presence of complexes.
Culture vesselSurf. area per well1Shared reagentsDNA transfectionRNAi transfection
Vol. of plating mediumVol. of dilution medium2DNALipofect- amine™
2000RNALipofect- amine™
2000
96-well0.3 cm2100 µl2 x 25 µl0.2 µg0.5 µl5 pmol0.25 µl
24-well2 cm2500 µl2 x 50 µl0.8 µg2.0 µl20 pmol1.0 µl
12-well4 cm21 ml2 x 100 µl1.6 µg4.0 µl40 pmol2.0 µl
6-well10 cm22 ml2 x 250 µl4.0 µg10 µl100 pmol5 µl
60-mm20 cm25 ml2 x 0.5 ml8.0 µg20 µl200 pmol10 µl
10-cm60 cm215 ml2 x 1.5 ml24 µg60 µl600 pmol30 µl
1 Surface areas may vary depending on the manufacturer.
2 Volumes of dilution medium in Step 2a & 2b of DNA or RNAi transfection protocols.
Purchaser Notification
This product is covered by one or more Limited Use Label Licenses (see the Invitrogen catalog or our web-site, www.invitrogen.com). By the use of this product you accept
the terms and conditions of all applicable Limited Use Label Licenses.
Limited Use Label License No. 27: Lipofectamine™ 2000 Reagent
Limited Use Label License No. 173: Inhibition of Gene Expression by Double-Stranded RNA Limited Use Label License No. 196: Stealth™ RNAi
©2000-2006 Invitrogen Corporation. All rights reserved.
Cat. No. 11668-027 Size: 0.75 ml Cat. No. 11668-019 Size: 1.5 ml Cat. No. 11668-500 Size: 15 ml
Store at +4°C (do not freeze)
Description
Lipofectamine™ 2000 is a proprietary formulation for the transfection of nucleic acids (DNA and RNA) into eukaryotic cells providing the following advantages:
? Highest transfection efficiency in many cell types and formats (e.g. 96-well). Refer to the Cell Lines database at www.invitrogen.com for a list of cell types successfully transfected.
? Nucleic acid-Lipofectamine™ 2000 complexes can be added directly to cells in culture medium, in the presence or absence of serum.
? It is not necessary to remove complexes or change/add medium after transfection, but complexes may be removed after 4-6 hours.
Important Guidelines for Transfection
? Use the procedure on page 2 to transfect cells with short interfering RNA
(siRNA) or Stealth™ RNAi.
? Use the procedure on page 3 to transfect cells with plasmid DNA.
? We recommend Opti-MEM® I Reduced Serum Medium (Cat. No. 31985-062)
to dilute Lipofectamine™ 2000 and nucleic acids before complexing.
? Do not add antibiotics to media during transfection as this causes cell death.
? Maintain the same seeding conditions between experiments.
? Test serum-free media for compatibility with Lipofectamine™ 2000 since some serum-free formulations (e.g. CD293, SFM II, VP-SFM) may inhibit cationic lipid-mediated transfection.
Note: For more tips for your RNAi experiment, refer to “Seven Steps to RNAi Success”. This manual is available from www.invitrogen.com/rnai or Technical Service, as are
cell-type specific RNAi transfection protocols (see “RNAi protocols”.)
Quality Control
Lipofectamine™ 2000 is tested for absence of microbial contamination with blood agar plates, Sabaraud dextrose agar plates, and fluid thioglycolate medium, and functionally by transfection of CHO-K1 cells with a reporter plasmid.
Part No.: 11668.2k.pps Rev. Date: 11 July 2006
For research use only. Not intended for any animal or human therapeutic or diagnostic use.
For technical support, contact tech_service@invitrogen.com.
Stealth™ RNAi or siRNA Transfection
Page 2
Use this brief procedure to transfect Stealth™ RNAi or siRNA into mammalian cells in a 24-well format. For other formats, see Scaling Up or Down Transfections (page 4). All amounts and volumes are given on a per well basis. Use this procedure as a starting point; optimize transfections as described in Optimizing Stealth™ RNAi or siRNA Transfection, especially if you are transfecting a mammalian cell line for the first time.
1. One day before transfection, plate cells in 500 µl of growth medium without antibiotics such that they will be 30-50% confluent at the time of transfection. Note: Transfecting cells at a lower density allows a longer interval between transfection and assay time, and minimizes the loss of cell viability due to cell overgrowth.
2. For each transfection sample, prepare oligomer-Lipofectamine™ 2000
complexes as follows:
a. Dilute 20 pmol Stealth™ RNAi or siRNA oligomer in 50 µl Opti-MEM® I Reduced Serum Medium without serum (final concentration of RNA when added to the cells is 33 nM). Mix gently.
b. Mix Lipofectamine™ 2000 gently before use, then dilute 1 µl in 50 µl Opti- MEM® I Reduced Serum Medium. Mix gently and incubate for 5 minutes at room temperature. Note: Proceed to Step c within 25 minutes.
c. After the 5-minute incubation, combine the diluted oligomer with the diluted Lipofectamine™ 2000. Mix gently and incubate for 20 minutes at room temperature (solution may appear cloudy).
3. Add the oligomer-Lipofectamine™ 2000 complexes to each well containing cells and medium. Mix gently by rocking the plate back and forth.
Incubate the cells at 37°C in a CO2 incubator for 24-96 hours until you are ready to assay for gene knockdown. Medium may be changed after 4-6 hours.
Optimizing Stealth™ RNAi or siRNA Transfection
To obtain the highest transfection efficiency and low non-specific effects, optimize transfection conditions by varying RNA and Lipofectamine™ 2000
concentrations. Test 10-50 pmol RNA and 0.5-1.5 µl Lipofectamine™ 2000 for 24- well format. Depending on the nature of the target gene, transfecting cells at higher densities may also be considered when optimizing conditions.
Plasmid DNA Transfection
Page 3
Use the following procedure to transfect DNA into mammalian cells in a 24-well format. For other formats, see Scaling Up or Down Transfections (page 4). All amounts and volumes are given on a per well basis. Prepare complexes using a
DNA (µg) to Lipofectamine™ 2000 (µl) ratio of 1:2 to 1:3 for most cell lines. Transfect cells at high cell density for high efficiency, high expression levels, and to minimize cytotoxicity. Optimization may be necessary (see Optimizing Plasmid DNA Transfection, page 4).
1. Adherent cells: One day before transfection, plate 0.5-2 x 105 cells in 500 µl of growth medium without antibiotics so that cells will be 90-95% confluent at the time of transfection.
Suspension cells: Just prior to preparing complexes, plate 4-8 x 105 cells in
500 µl of growth medium without antibiotics.
2. For each transfection sample, prepare complexes as follows:
a. Dilute DNA in 50 µl of Opti-MEM® I Reduced Serum Medium without serum (or other medium without serum). Mix gently.
b. Mix Lipofectamine™ 2000 gently before use, then dilute the appropriate amount in 50 µl of Opti-MEM® I Medium. Incubate for 5 minutes at room temperature. Note: Proceed to Step c within 25 minutes.
c. After the 5 minute incubation, combine the diluted DNA with diluted
Lipofectamine™ 2000 (total volume = 100 µl). Mix gently and incubate for
20 minutes at room temperature (solution may appear cloudy). Note:
Complexes are stable for 6 hours at room temperature.
3. Add the 100 µl of complexes to each well containing cells and medium. Mix gently by rocking the plate back and forth.
4. Incubate cells at 37°C in a CO2 incubator for 18-48 hours prior to testing for transgene expression. Medium may be changed after 4-6 hours.
5. For stable cell lines: Passage cells at a 1:10 (or higher dilution) into fresh growth medium 24 hours after transfection. Add selective medium (if desired) the following day.
Optimizing Plasmid DNA Transfection
To obtain the highest transfection efficiency and low cytotoxicity, optimize transfection conditions by varying cell density as well as DNA and
Page 4
Lipofectamine™ 2000 concentrations. Make sure that cells are greater than 90%
confluent and vary DNA (µg): Lipofectamine™ 2000 (µl) ratios from 1:0.5 to 1:5.
Scaling Up or Down Transfections
To transfect cells in different tissue culture formats, vary the amounts of Lipofectamine™ 2000, nucleic acid, cells, and medium used in proportion to the relative surface area, as shown in the table. With automated, high-throughput systems, a complexing volume of 50 µl is recommended for transfections in 96- well plates. Note: You may perform rapid 96-well plate transfections by plating cells directly into the transfection mix. Prepare complexes in the plate and directly add cells at twice the cell density as in the basic protocol in a 100 µl volume. Cells will adhere as usual in the presence of complexes.
Culture vesselSurf. area per well1Shared reagentsDNA transfectionRNAi transfection
Vol. of plating mediumVol. of dilution medium2DNALipofect- amine™
2000RNALipofect- amine™
2000
96-well0.3 cm2100 µl2 x 25 µl0.2 µg0.5 µl5 pmol0.25 µl
24-well2 cm2500 µl2 x 50 µl0.8 µg2.0 µl20 pmol1.0 µl
12-well4 cm21 ml2 x 100 µl1.6 µg4.0 µl40 pmol2.0 µl
6-well10 cm22 ml2 x 250 µl4.0 µg10 µl100 pmol5 µl
60-mm20 cm25 ml2 x 0.5 ml8.0 µg20 µl200 pmol10 µl
10-cm60 cm215 ml2 x 1.5 ml24 µg60 µl600 pmol30 µl
1 Surface areas may vary depending on the manufacturer.
2 Volumes of dilution medium in Step 2a & 2b of DNA or RNAi transfection protocols.
Purchaser Notification
This product is covered by one or more Limited Use Label Licenses (see the Invitrogen catalog or our web-site, www.invitrogen.com). By the use of this product you accept
the terms and conditions of all applicable Limited Use Label Licenses.
Limited Use Label License No. 27: Lipofectamine™ 2000 Reagent
Limited Use Label License No. 173: Inhibition of Gene Expression by Double-Stranded RNA Limited Use Label License No. 196: Stealth™ RNAi
©2000-2006 Invitrogen Corporation. All rights reserved.
描述
风险提示：丁香通仅作为第三方平台，为商家信息发布提供平台空间。用户咨询产品时请注意保护个人信息及财产安全，合理判断，谨慎选购商品，商家和用户对交易行为负责。对于医疗器械类产品，请先查证核实企业经营资质和医疗器械产品注册证情况。
- 作者
- 内容
- 询问日期
文献和实验相关实验
lipofectamine 2000脂质体转染293ft细胞经验
相关专题 293细胞 总有一种转染方法适合你 问： 我用lipofectamine 2000转染293ft细胞，细胞老是死亡，是什么原因?求各位先知不吝赐教。我的protocol如下： Day 0：细胞传代后计数、seed 2.0x105Cell s and 1.0x106 cells per well of plate Day 1：细胞达到70%汇合后准备:溶液1：200ul 无血清培养基 + 4 ul
lipofectamine 2000脂质体转染293ft细胞经验
相关专题 293细胞 总有一种转染方法适合你 问： 我用lipofectamine 2000转染 293ft细胞，细胞老是死亡，是什么原因?求各位先知不吝赐教。我的protocol如下： Day 0：细胞传代后计数、seed 2.0x105Cell s and 1.0x106 cells per well of plate Day 1：细胞达到70%汇合后准备:溶液1：200ul 无血清培养基 + 4 ul
使用Lipofectamine™2000转染Stealth™ RNA或者siRNA进入哺乳动物细胞
前言 Lipofectamine™ 2000试剂是一项专利配方，用于高效转染Stealth™ RNA或者短的干扰RNA（siRNA）到哺乳动物细胞，以进行RNAi分析（1，2）。该说明书提供了一般的指导以及使用Lipofectamine™ 2000转染Stealth™ RNA或者siRNAj进入哺乳动物细胞的步骤。提供推荐的起始使用试剂剂量。为了获得最佳的RNAi实验结果，需要针对哺乳动物细胞系和目的
技术资料暂无技术资料 索取技术资料
脂质体2000，Lip2000，Lipofectamine™ 2000 Transfection Reagent
￥2600
