Human BCAP31 (BAP31) knockout HEK-293T cell line (ab266634)
Overview
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Product name
Human BCAP31 (BAP31) knockout HEK-293T cell line -
Parental Cell Line
HEK293T -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 2 -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
Tested applications
Suitable for: WBmore details -
Biosafety level
2 -
General notes
Recommended control: Human wild-type HEK293T cell line (ab255449). 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 -
STR Analysis
Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12 -
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
May play a role in anterograde transport of membrane proteins from the endoplasmic reticulum to the Golgi. May be involved in CASP8-mediated apoptosis. -
Tissue specificity
Ubiquitous. -
Involvement in disease
Note=BCAP31 is deleted in the chromosome Xq28 deletion syndrome which involves BCAP31 and the and the promoter region of ABCD1. -
Sequence similarities
Belongs to the BCAP29/BCAP31 family. -
Post-translational
modificationsCleaved by CASP8 and other caspases. -
Cellular localization
Endoplasmic reticulum membrane. Endoplasmic reticulum-Golgi intermediate compartment membrane. May shuttle between the ER and the intermediate compartment/cis-Golgi complex. - Information by UniProt
Associated products
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KO cell lysates
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab266634 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: 28 kDa.
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Notes |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 28 kDa. |
Images
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All lanes : Anti-BAP31 antibody [EPR3878(2)] (ab109304) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : BCAP31 knockout HEK293T cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : NCCIT cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 28 kDa
Observed band size: 28 kDaLanes 1-4: Merged signal (red and green). Green - ab109304 observed at 28 kDa. Red - loading control ab7291 observed at 50 kDa.
ab109304 Anti-BAP31 antibody [EPR3878(2)] was shown to specifically react with BAP31 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266634 (knockout cell lysate ab257857) was used. Wild-type and BAP31 knockout samples were subjected to SDS-PAGE. ab109304 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-BAP31 antibody [7A3BB6] (ab112993) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : BCAP31 knockout HEK293T cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : NCCIT cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution
Predicted band size: 28 kDa
Observed band size: 28 kDaLanes 1-4: Merged signal (red and green). Green - ab112993 observed at 28 kDa. Red - loading control ab52901 observed at kDa.
ab112993 Anti-BAP31 antibody [7A3BB6] was shown to specifically react with BAP31 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266634 (knockout cell lysate ab257857) was used. Wild-type and BAP31 knockout samples were subjected to SDS-PAGE. ab112993 and Anti-beta Tubulin [EP1331Y] - Microtubule Marker (ab52901) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Homozygous: Insertion of the selection cassette in exon 2
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Representative images of BCAP31 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab266634 has not yet been referenced specifically in any publications.