Mouse CCL7 knockout RAW 264.7 cell lysate (ab275525)
Overview
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Product name
Mouse CCL7 knockout RAW 264.7 cell lysate
See all MCP3 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9.
Treatments:
Mouse CCL7 knockout RAW 264.7 cell lysate - Untreated
Mouse wild-type RAW 264.7 cell lysate - Untreated
Mouse CCL7 knockout RAW 264.7 cell lysate - PMA (80 nM, 24h), LPS (100 ng/ml, 6h) and Brefeldin A (5 µg/ml, in the last 5h of LPS treatment)
Mouse wild-type RAW 264.7 cell lysate - PMA (80 nM, 24h), LPS (100 ng/ml, 6h) and Brefeldin A (5 µg/ml, in the last 5h of LPS treatment) -
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
Knockout validation
Sanger Sequencing, Western Blot (WB)Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
Notes
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.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.
Tested applications
Suitable for: WBmore detailsProperties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab277357 - Mouse CCL7 knockout RAW 264.7 cell lysate - PMA, LPS + Brefeldin A treated 1 x 100µg ab277356 - Mouse CCL7 knockout RAW 264.7 cell lysate 1 x 100µg ab277385 - Mouse wild-type RAW 264.7 cell lysate - PMA, LPS + Brefeldin A treated 1 x 100µg ab277354 - Mouse wild-type RAW 264.7 cell lysate 1 x 100µg -
Research areas
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Cell type
leukaemic monocyte macrophage -
Disease
Carcinoma -
Gender
Male
Target
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Function
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. -
Sequence similarities
Belongs to the intercrine beta (chemokine CC) family. -
Post-translational
modificationsO-glycosylated. -
Cellular localization
Secreted. - Information by UniProt
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Alternative names
- C-C motif chemokine 7
- Ccl7
- CCL7_HUMAN
see all
Properties
-
Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab277357 - Mouse CCL7 knockout RAW 264.7 cell lysate - PMA, LPS + Brefeldin A treated 1 x 100µg ab277356 - Mouse CCL7 knockout RAW 264.7 cell lysate 1 x 100µg ab277385 - Mouse wild-type RAW 264.7 cell lysate - PMA, LPS + Brefeldin A treated 1 x 100µg ab277354 - Mouse wild-type RAW 264.7 cell lysate 1 x 100µg -
Research areas
-
Cell type
leukaemic monocyte macrophage -
Disease
Carcinoma -
Gender
Male
Images
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Western blot - Mouse CCL7 knockout RAW 264.7 cell lysate (ab275525)
Lane 1: Wild-type RAW 264.7 untreated control cell lysate 30 ug
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 ug/ml, 5 h) cell lysate 30 ug
Lane 3: CCL7/MCP3 knockout RAW 264.7 untreated cell lysate 30 ug
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 ug/ml, 5 h) cell lysate 30 ug
Lane 5: J774A.1 Glucose treated (138.8 mMol/L, 8 h) plus Brefeldin A treated (5 ug/ml, 6 h) and Brefeldin A (ab120299) treated (5 ug/ml, 5 h) cell lysate 30 ug
Lane 6: J774A.1 Glucose treated (5.6 mMol/L, 8 h) plus Brefeldin A treated (5 ug/ml, 6 h) and Brefeldin A (ab120299) treated (5 ug/ml, 5 h) cell lysate 30 ug
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 37kDa.
ab228979 was shown to react with MCP3 in wild-type RAW 264.7 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. -
Sanger Sequencing - Human CCL7 knockout RAW264.7 cell lysate