Human LCN2 knockout SW480 cell lysate (ab270509)
Overview
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Product name
Human LCN2 knockout SW480 cell lysate
See all Lipocalin-2 / NGAL kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
SW480 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 1 bp deletion, 4 bp deletion; Frameshift: 99.97% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), 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.
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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.Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.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.
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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab280562 - Human LCN2 knockout SW480 cell lysate 1 x 100µg ab269601 - Human wild-type SW480 cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Male
Target
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Function
Iron-trafficking protein involved in multiple processes such as apoptosis, innate immunity and renal development. Binds iron through association with 2,5-dihydroxybenzoic acid (2,5-DHBA), a siderophore that shares structural similarities with bacterial enterobactin, and delivers or removes iron from the cell, depending on the context. Iron-bound form (holo-24p3) is internalized following binding to the SLC22A17 (24p3R) receptor, leading to release of iron and subsequent increase of intracellular iron concentration. In contrast, association of the iron-free form (apo-24p3) with the SLC22A17 (24p3R) receptor is followed by association with an intracellular siderophore, iron chelation and iron transfer to the extracellular medium, thereby reducing intracellular iron concentration. Involved in apoptosis due to interleukin-3 (IL3) deprivation: iron-loaded form increases intracellular iron concentration without promoting apoptosis, while iron-free form decreases intracellular iron levels, inducing expression of the proapoptotic protein BCL2L11/BIM, resulting in apoptosis. Involved in innate immunity, possibly by sequestrating iron, leading to limit bacterial growth. -
Tissue specificity
Expressed in bone marrow and in tissues that are prone to exposure to microorganism. High expression is found in bone marrow as well as in uterus, prostate, salivary gland, stomach, appendix, colon, trachea and lung. Not found in the small intestine or peripheral blood leukocytes. -
Sequence similarities
Belongs to the calycin superfamily. Lipocalin family. -
Cellular localization
Secreted. Upon binding to the SLC22A17 (24p3R) receptor, it is internalized. - Information by UniProt
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Alternative names
- 24p3
- 25 kDa alpha 2 microglobulin related subunit of MMP9
- 25 kDa alpha-2-microglobulin-related subunit of MMP-9
see all
Associated products
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KO cell lines
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Related Products
- Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075)
- Anti-Lipocalin-2 / NGAL antibody [EPR5084] - BSA and Azide free (ab187969)
- Anti-Lipocalin-2 / NGAL antibody [14] (ab188551)
- Anti-Lipocalin-2 / NGAL antibody [EPR19912] (ab206427)
- Anti-Lipocalin-2 / NGAL antibody [EPR19912] - Low endotoxin, Azide free (ab224264)
- Anti-Lipocalin-2 / NGAL antibody [5G5] (ab23477)
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab270509 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
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| WB |
Use at an assay dependent concentration. Predicted molecular weight: 22 kDa.
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| Notes |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 22 kDa. |
Images
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Next Generation Sequencing - Human LCN2 knockout SW480 cell lysate (ab270509)Knockout achieved by CRISPR/Cas9; X = 1 bp deletion, 4 bp deletion; Frameshift: 99.97%
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Western blot - Human LCN2 knockout SW480 cell lysateLane 1: Wild-type SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate, 20 ug
Lane 2: LCN2 knockout SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate, 20 ug
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate, 20 ug
Lane 4: SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate, 20 ugLanes 1 - 4: Merged signal (red and green). Green - ab206427 observed at 25 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab206427 was shown to react with Lipocalin-2 / NGAL in wild-type SW480 cells in Western blot with loss of signal observed in LCN2 knockout cell line ab270486 (knockout cell lysate ab270509). Wild-type SW480 and LCN2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab206427 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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Western blot - Human LCN2 knockout SW480 cell lysateLane 1: Wild-type SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate, 20 ug
Lane 2: LCN2 knockout SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate, 20 ug
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate, 20 ug
Lane 4: SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate, 20 ugLanes 1 - 4: Merged signal (red and green). Green - ab188551 observed at 24 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.
ab188551 was shown to react with Lipocalin-2 / NGAL in wild-type SW480 cells in Western blot with loss of signal observed in LCN2 knockout cell line ab270486 (knockout cell lysate ab270509). Wild-type SW480 and LCN2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab188551 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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Western blot - Human LCN2 knockout SW480 cell lysateLane 1: Wild-type SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate, 20 ug
Lane 2: LCN2 knockout SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate, 20 ug
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate, 20 ug
Lane 4: SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate, 20 ugLanes 1 - 4: Merged signal (red and green). Green - ab125075 observed at 23 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab125075 was shown to react with Lipocalin-2 / NGAL in wild-type SW480 cells in Western blot with loss of signal observed in LCN2 knockout cell line ab270486 (knockout cell lysate ab270509). Wild-type SW480 and LCN2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab125075 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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Western blot - Human LCN2 knockout SW480 cell lysateLane 1: Wild-type SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate, 20 ug
Lane 2: LCN2 knockout SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate, 20 ug
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate, 20 ug
Lane 4: SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate, 20 ugLanes 1 - 4: Merged signal (red and green). Green - ab23477 observed at 23 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.
ab23477 was shown to react with Lipocalin-2 / NGAL in wild-type SW480 cells in Western blot with loss of signal observed in LCN2 knockout knockout cell line ab270486 (knockout cell lysate ab270509). Wild-type SW480 and LCN2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab23477 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab270509 has not yet been referenced specifically in any publications.