Human DNAJB1 (Hsp40) knockout HEK-293T cell line
- Advanced Validation
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DNAJB1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 29 bp deletion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
DNAJ 1, DNAJ B1, DNAJB1 protein, DNJB1_HUMAN, DnaJ (Hsp40) homolog subfmaily B member 1, DnaJ heat shock protein family (Hsp40) member B1, DnaJ homolog subfamily B member 1, DnaJ protein homolog 1, HDJ-1, HSPF 1, Heat shock 40 kDa protein 1, Heat shock 40kD protein 1, Heat shock protein 40, Hsp 40, Human DnaJ protein 1, RSPH16B, Radial spoke 16 homolog B, Sis1
- WB
Lab
Western blot - Human DNAJB1 (Hsp40) knockout HEK-293T cell line (AB266344)
Lanes 1-4 : Merged signal (red and green). Green - ab223607 observed at 39 kDa. Red - loading control ab52901 observed at kDa.
ab223607 Anti-Hsp40 antibody [3B9.E6] was shown to specifically react with Hsp40 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266344 (knockout cell lysate ab258400) was used. Wild-type and Hsp40 knockout samples were subjected to SDS-PAGE. ab223607 and Anti-beta Tubulin [EP1331Y] - Microtubule Marker (ab52901) were incubated overnight at 4°C at 1 in 500 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.
All lanes:
Western blot - Anti-Hsp40 antibody [3B9.E6] (<a href='/en-us/products/primary-antibodies/hsp40-antibody-3b9e6-ab223607'>ab223607</a>) at 1/500 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
DNAJB1 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human DNAJB1 (Hsp40) knockout HEK-293T cell line (ab266344)
Lane 3:
A549 cell lysate at 20 µg
Lane 4:
Daudi cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution
Predicted band size: 38 kDa
Observed band size: 39 kDa
false
- Cell Culture
Unknown
Cell Culture - Human DNAJB1 (Hsp40) knockout HEK-293T cell line (AB266344)
Representative images of DNAJB1 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.
- Sanger seq
Unknown
Sanger Sequencing - Human DNAJB1 (Hsp40) knockout HEK-293T cell line (AB266344)
Homozygous : 29 bp deletion in exon 1
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Human DNAJB1 (Hsp40) knockout HEK-293T cell line (AB266344)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized DNAJB1 (Hsp40) KO HEK293T ( DNAJB1 (Hsp40) knockout human embryonic kidney epithelial cell) (ab266344) cells labelling Hsp40 with ab323655 at 1/50 (10.0 ug/ml) dilution (Green).
Confocal image showing nuclear and cytoplasmic staining in parental HEK293T cells and no staining in DNAJB1 (Hsp40) KO HEK293T cells(shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/ab195889 200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
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 seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate 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. 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.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Hsp40 proteins participate in cell signaling and stress response by modulating the activity of Hsp70 forming a functional complex with it to guide protein folding processes. This process is essential for protein quality control and cellular protection. It interacts with unfolded proteins and delivers them to Hsp70 enhancing its ATPase activity necessary for the proper functioning of these molecular chaperones. The expression of Hsp40 in stress conditions highlights its importance in the protein quality control system.
Pathways
Hsp40 is involved in important cellular mechanisms like the unfolded protein response and protein catabolism pathways. It works closely with Hsp70 playing a pivotal role in managing protein misfolding and aggregation vital for cellular stress response and recovery. In these pathways Hsp40's function ensures the proper folding and degradation of damaged proteins maintaining proteostasis in the cell.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Cell Lines & Lysates
AB266343
Human RAD23A (hHR23A) knockout HEK-293T cell line
Advanced Validation
- 0
- 0
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com