Human ACP1 (Acid phosphatase) knockout HEK-293 cell line (ab261859)
Overview
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Product name
Human ACP1 (Acid phosphatase) knockout HEK-293 cell line -
Parental Cell Line
HEK-293 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 100% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), Western Blot (WB) -
Tested applications
Suitable for: WBmore details -
Biosafety level
2 -
General notes
Recommended control: Human wild-type HEK-293 cell line (ab259776). 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 (High Glucose) + 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 bath for 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 2x104 cells/cm2. 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 2x104 cells/cm2 is recommended.
- A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.
- Cells should be passaged when they have achieved 80-90% confluence.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
We will provide viable cells that proliferate on revival.
Properties
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Number of cells
1 x 106 cells/vial, 1 mL -
Adherent /Suspension
Adherent -
Tissue
Kidney -
Cell type
epithelial -
Gender
Female -
Mycoplasma free
Yes -
Storage instructions
Shipped on Dry Ice. Store in liquid nitrogen. -
Storage buffer
Constituents: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether -
Research areas
Target
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Function
Acts on tyrosine phosphorylated proteins, low-MW aryl phosphates and natural and synthetic acyl phosphates. Isoform 3 does not possess phosphatase activity. -
Tissue specificity
T-lymphocytes express only isoform 2. -
Sequence similarities
Belongs to the low molecular weight phosphotyrosine protein phosphatase family. -
Cellular localization
Cytoplasm. - Information by UniProt
Associated products
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KO cell lysates
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Related Products
- Anti-Acid phosphatase/ACP1 antibody [EPR9839] (ab166896)
- Anti-Acid phosphatase antibody [EPR21787] (ab235448)
- Anti-Acid phosphatase antibody [EPR21791] (ab235449)
- Anti-Acid phosphatase antibody [EPR21787] - BSA and Azide free (ab238888)
- Anti-Acid phosphatase antibody [EPR21791] - BSA and Azide free (ab238913)
- Anti-Acid phosphatase/ACP1 antibody [EPR9839] - BSA and Azide free (ab249379)
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab261859 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.
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Notes |
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WB
Use at an assay dependent concentration. |
Images
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All lanes : Anti-Acid phosphatase antibody [EPR21791] (ab235449) at 1/1000 dilution
Lane 1 : Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : ACP1 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : K562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate
Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 18 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab235449 observed at 18 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab235449 was shown to react with Acid phosphatase in wild-type HEK-293 cells in western blot with loss of signal observed in ACP1 knockout cell line ab261859 (knockout cell lysate ab261668). Wild-type and ACP1 knockout HEK-293 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab235449 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 hour at room temperature before imaging.
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All lanes : Anti-Acid phosphatase/ACP1 antibody [EPR9839] (ab166896) at 1/1000 dilution
Lane 1 : Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : ACP1 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : K562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate
Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 18 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab166896 observed at 18 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab166896 was shown to recognize ACP1 (Acid phosphatase 1) in wild-type HEK-293 cells as signal was lost at the expected MW in ACP1 knockout cell line ab261859 (knockout cell lysate ab261668). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and ACP1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. Ab166896 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-Acid phosphatase/ACP1 antibody [EPR9838(2)] (ab180524) at 1/1000 dilution
Lane 1 : Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : ACP1 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : K562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate
Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 18 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab180524 observed at 18 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab180524 was shown to specifically react with ACP1 (Acid phosphatase 1) in wild-type HEK-293 cells as signal was lost in ACP1 knockout cell line ab261859 (knockout cell lysate ab261668). Wild-type and ACP1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. Ab180524 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-Acid phosphatase antibody [EPR21787] (ab235448) at 1/1000 dilution
Lane 1 : Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : ACP1 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : K562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate
Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.Lanes 1 - 4: Merged signal (red and green). Green - ab235448 observed at 18 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab235448 was shown to specifically react with ACP1 (Acid phosphatase 1) in wild-type HEK-293 cells as signal was lost in ACP1 knockout cell line ab261859 (knockout cell lysate ab261668). Wild-type and ACP1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. Ab235448 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.
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X = 1 bp insertion
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab261859 has not yet been referenced specifically in any publications.