小鼠CCL7 knockout RAW 264.7 cell line (ab273755)
概述
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产品名称
小鼠CCL7 knockout RAW 264.7 cell line -
Parental Cell Line
RAW 264.7 -
Organism
Mouse -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 85 bp deletion in exon 2 -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
经测试应用
适用于: WB, Flow Cytmore details -
Biosafety level
2 -
常规说明
Recommended control: Mouse wild-type RAW 264.7 cell line (ab275474). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: DMEM + 10% FBS
Initial handling guidelines: Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water for bath approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily.Subculture guidelines:
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x105 cells/mL is recommended.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
We will provide viable cells that proliferate on revival.
性能
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Number of cells
1 x 106 cells/vial, 1 mL -
Adherent /Suspension
Adherent -
Tissue
Lymphatic -
Cell type
leukaemic monocyte macrophage -
Disease
Carcinoma -
Gender
Male -
Mycoplasma free
Yes -
存放说明
Shipped on Dry Ice. Store in liquid nitrogen. -
存储溶液
Constituents: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether -
研究领域
靶标
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功能
Chemotactic factor that attracts monocytes and eosinophils, but not neutrophils. Augments monocyte anti-tumor activity. Also induces the release of gelatinase B. This protein can bind heparin. Binds to CCR1, CCR2 and CCR3. -
序列相似性
Belongs to the intercrine beta (chemokine CC) family. -
翻译后修饰
O-glycosylated. -
细胞定位
Secreted. - Information by UniProt
相关产品
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Biochemicals
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KO cell lysates
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab273755于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 11 kDa.
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Flow Cyt |
Use at an assay dependent concentration.
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说明 |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 11 kDa. |
Flow Cyt
Use at an assay dependent concentration. |
图片
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All lanes : Anti-MCP3 antibody [EPR22649-155] (ab228979) at 1/1000 dilution
Lane 1 : Wild-type RAW 264.7 untreated control cell lysate
Lane 2 : Wild-type RAW 264.7 PMA treated (80 nM, 24 h) plus LPS treated (100 ng/ml, 6 h) and Brefeldin A (ab120299) treated (5 µg/ml, 5 h) cell lysate
Lane 3 : CCL7/MCP3 knockout RAW 264.7 untreated cell lysate
Lane 4 : CCL7/MCP3 knockout RAW 264.7 PMA treated (80 nM, 24 h) plus LPS treated (100 ng/ml, 6 h) and Brefeldin A (ab120299) treated (5 µg/ml, 5 h) cell lysate
Lane 5 : J774A.1 Glucose treated (138.8 mMol/L, 8 h) plus Brefeldin A treated (5 µg/ml, 6 h) and Brefeldin A (ab120299) treated (5 µg/ml, 5 h) cell lysate
Lane 6 : J774A.1 Glucose treated (5.6 mMol/L, 8 h) plus Brefeldin A treated (5 µg/ml, 6 h) and Brefeldin A (ab120299) treated (5 µg/ml, 5 h) cell lysate
Lysates/proteins at 30 µg per lane.
Performed under reducing conditions.
Predicted band size: 11 kDa
Observed band size: 14 kDa why is the actual band size different from the predicted?Lanes 1 - 6: Merged signal (red and green). Green - ab228979 observed at 14 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab228979 was shown to react with MCP3 in RAW 264.7 wild-type cells in Western blot with loss of signal observed in CCL7 knockout sample. Wild-type and CCL7 knockout RAW 264.7 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab228979 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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Homozygous: 85 bp deletion in exon 2
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (0)
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