Human ANP32B (PHAPI2/APRIL) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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ANP32B KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 13 bp deletion in exon 1.
View Alternative Names
AN32B_HUMAN, ANP 32B, APRIL, Acidic (leucine rich) nuclear phosphoprotein 32 family member B, Acidic leucine-rich nuclear phosphoprotein 32 family member B, Acidic nuclear phosphoprotein 32 family member B, Acidic protein rich in leucines, OTTHUMP0000006377, PAL 31, PHAPI 2, PHAPI2 protein, PHAPI2a, Proliferation related acidic leucine rich protein, Putative HLA-DR-associated protein I-2, Silver stainable protein SSP 29, Silver-stainable protein SSP29, Ssp 29
- WB
Lab
Western blot - Human ANP32B (PHAPI2/APRIL) knockout HEK-293T cell line (AB266818)
Lanes 1-4 : Merged signal (red and green). Green - ab200836 observed at 29 kDa. Red - loading control ab8245 observed at 36 kDa.
ab200836 Anti-PHAPI2 / APRIL antibody [EPR14588] was shown to specifically react with PHAPI2/ APRIL in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266818 (knockout cell lysate ab257831) was used. Wild-type and PHAPI2 / APRIL knockout samples were subjected to SDS-PAGE. ab200836 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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.
All lanes:
Western blot - Anti-PHAPI2 / APRIL antibody [EPR14588] (<a href='/en-us/products/primary-antibodies/phapi2-april-antibody-epr14588-ab200836'>ab200836</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human ANP32B (PHAPI2/APRIL) knockout HEK-293T cell line (ab266818)
Lane 2:
ANP32B knockout HEK293T cell lysate at 20 µg
Lane 3:
Jurkat cell lysate at 20 µg
Lane 4:
LNCaP cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 28 kDa
Observed band size: 29 kDa
false
- Cell Culture
Lab
Cell Culture - Human ANP32B (PHAPI2/APRIL) knockout HEK-293T cell line (AB266818)
Representative images ANP32B 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 M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human ANP32B (PHAPI2/APRIL) knockout HEK-293T cell line (AB266818)
Homozygous : 13 bp deletion in exon1
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 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
PHAPI2 regulates immune system processes by playing a role in B cell development and activity. It functions as part of larger protein complexes binding with proteins such as BCMA to influence signaling pathways. The interactions within these complexes help modulate processes critical for proper immune response and maintenance of immune system homeostasis.
Pathways
PHAPI2 is deeply involved in both the NF-kB signaling pathway and the PI3K/AKT pathway. These pathways are essential for cellular growth survival and proliferation. Through its involvement PHAPI2 also connects with proteins like NF-kB and AKT1 which are key regulatory proteins within these pathways. This connection enables PHAPI2 to act as a node integrating signals for various cellular responses.
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
AB200836
Anti-PHAPI2 / APRIL antibody [EPR14588]
RabMAb|Recombinant|KO Validated
![Immunocytochemistry/ Immunofluorescence - Anti-PHAPI2 / APRIL antibody [EPR14588] (AB200836)](https://content.abcam.com/products/images/phapi2-april-antibody-epr14588-ab200836--immunocytochemistry-immunofluorescence-img14994.jpg)
- 0
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![Western blot - Anti-BAFF antibody [EPR22238] (AB224710)](https://content.abcam.com/products/images/baff-antibody-epr22238-ab224710--western-blot-img54560.jpg)
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