Human HDAC2 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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HDAC2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 2 and 1 bp insertion in exon 2.
View Alternative Names
D10Wsu179e, HD 2, HDAC2_HUMAN, Histone deacetylase 2, Histone deacetylase 2 (HD2), OTTHUMP00000017046, OTTHUMP00000227077, OTTHUMP00000227078, RPD 3, YAF1, YY1 associated factor 1, YY1 transcription factor binding protein, Yy1bp, transcriptional regulator homolog RPD3
- IHC-P
Supplier Data
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Human HDAC2 knockout HEK-293T cell line (AB266590)
Immunohistochemical analysis of paraffin-embedded fixed (A) Wild-type HEK293T (human embryonic kidney epithelial cell) cell pellet. (B) HDAC2 knockout HEK293T (ab266590) cell pellet staining HDAC2 with ab32117 at 1/10000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Counter-staining used was hematoxylin. Positive staining on (A) Wild-type HEK293T cell pellet, no staining on HDAC2 knockout HEK293T (ab266590) cell pellet. The section was incubated with ab32117 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Human HDAC2 knockout HEK-293T cell line (AB266590)
Immunocytochemistry/ Immunofluorescence analysis of Wild-type HEK293T/HDAC2 KO HEK293T (HDAC2 knockout human embryonic kidney epithelial cell) (ab266590) cells labeling HDAC2 with ab32117 at 1/200 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody at 1/1000 dilution (2 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (2.5 µg/ml). DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control. Confocal image showing nuclear staining in Parental HEK293T cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
- WB
Lab
Western blot - Human HDAC2 knockout HEK-293T cell line (AB266590)
Lanes 1-2 : Merged signal (red and green). Green - ab124974 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.
ab124974 Anti-HDAC2 antibody [EPR5001] was shown to specifically react with HDAC2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266590 (knockout cell lysate ab256939) was used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab124974 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 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.
All lanes:
Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (<a href='/en-us/products/primary-antibodies/hdac2-antibody-epr5001-chip-grade-ab124974'>ab124974</a>) at 1/10000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
HDAC2 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human HDAC2 knockout HEK-293T cell line (ab266590)
Predicted band size: 40 kDa,48 kDa,55 kDa,59 kDa
Observed band size: 40 kDa,48 kDa,59 kDa,60 kDa
false
- WB
Unknown
Western blot - Human HDAC2 knockout HEK-293T cell line (AB266590)
Lanes 1-2 : Merged signal (red and green). Green - ab219053 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.
ab219053 Anti-HDAC2 antibody [EPR20117] was shown to specifically react with HDAC2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266590 (knockout cell lysate ab256939) was used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab219053 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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.
All lanes:
Western blot - Anti-HDAC2 antibody [EPR20117] (<a href='/en-us/products/primary-antibodies/hdac2-antibody-epr20117-ab219053'>ab219053</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
HDAC2 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human HDAC2 knockout HEK-293T cell line (ab266590)
Predicted band size: 55 kDa
Observed band size: 60 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human HDAC2 knockout HEK-293T cell line (AB266590)
Allele-2 : 1 bp insertion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human HDAC2 knockout HEK-293T cell line (AB266590)
Allele-1 : 11 bp deletion in exon2
- Cell Culture
Unknown
Cell Culture - Human HDAC2 knockout HEK-293T cell line (AB266590)
Representative images of HDAC2 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 a EVOS XL Core microscope.
Reactivity data
Product details
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.
We will provide viable cells that proliferate on revival.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
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
HDAC2 is involved in cell cycle regulation and differentiation. It is part of the co-repressor complex and interacts with other proteins such as Sin3 and NuRD to repress transcription. These interactions are vital for maintaining normal cellular functions and ensuring proper response to external signals. This protein is also connected to HEK293T cells which are a common model in scientific studies due to their robust expression of proteins like HDAC2.
Pathways
HDAC2 participates in the regulation of transcriptional activity and cellular stress response. It is an important player in the histone modification pathway interacting with proteins such as HDAC1 and transcriptional regulators like REST. HDAC2 is also linked with the Notch signaling pathway which is essential for cell fate decisions. These pathways highlight its role in both maintaining cellular homeostasis and adapting to changes in the cellular environment.
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
Primary Antibodies
AB32117
BOND RX™ Validated|RabMAb|Advanced Validation|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [Y461] (AB32117)](https://content.abcam.com/products/images/hdac2-antibody-y461-ab32117--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img178012.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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