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AB266142

Human HNRNPUL1 (E1B-AP5) knockout HEK-293T cell line

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HNRNPUL1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon 3 and Insertion of the selection cassette in exon 3. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
4 Images
Western blot - Human HNRNPUL1 (E1B-AP5) knockout HEK-293T cell line (AB266142)
  • WB

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Western blot - Human HNRNPUL1 (E1B-AP5) knockout HEK-293T cell line (AB266142)

Lanes 1-3 : Merged signal (red and green). Green - ab68480 observed at 96 kDa. Red - loading control ab8245 observed at 36 kDa.

ab68480 Anti-E1B-AP5 antibody [EP2633Y] was shown to specifically react with E1B-AP5 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266142 (knockout cell lysate ab257993) was used. Wild-type and E1B-AP5 knockout samples were subjected to SDS-PAGE. ab68480 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-E1B-AP5 antibody [EP2633Y] (<a href='/en-us/products/primary-antibodies/e1b-ap5-antibody-ep2633y-ab68480'>ab68480</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

HNRNPUL1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human HNRNPUL1 (E1B-AP5) knockout HEK-293T cell line (ab266142)

Lane 3:

Raji 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: 96 kDa

Observed band size: 96 kDa

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Sanger Sequencing - Human HNRNPUL1 (E1B-AP5) knockout HEK-293T cell line (AB266142)
  • Sanger seq

Unknown

Sanger Sequencing - Human HNRNPUL1 (E1B-AP5) knockout HEK-293T cell line (AB266142)

Allele-1 : Insertion of the selection cassette in exon 3

Sanger Sequencing - Human HNRNPUL1 (E1B-AP5) knockout HEK-293T cell line (AB266142)
  • Sanger seq

Unknown

Sanger Sequencing - Human HNRNPUL1 (E1B-AP5) knockout HEK-293T cell line (AB266142)

Allele-2 : 16 bp deletion in exon 3.

Cell Culture - Human HNRNPUL1 (E1B-AP5) knockout HEK-293T cell line (AB266142)
  • Cell Culture

Lab

Cell Culture - Human HNRNPUL1 (E1B-AP5) knockout HEK-293T cell line (AB266142)

Representative images HNRNPUL1 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.

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon 3 and Insertion of the selection cassette in exon 3

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Complementary Products

Primary Antibodies

AB68480

Anti-E1B-AP5 antibody [EP2633Y]

RabMAb|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E1B-AP5 antibody [EP2633Y] (AB68480)

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Proteins & Peptides

AB313361

Recombinant Human WFDC2 Protein (His tag)

Mass Spectrometry - Recombinant Human WFDC2 Protein (His tag) (AB313361)

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Proteins & Peptides

AB217407

Recombinant human Vitronectin/S-Protein (Animal Free)

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Proteins & Peptides

AB95933

Recombinant human XIAP protein

Functional Studies - Recombinant human XIAP protein (AB95933)

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  • 0
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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

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.

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What's included?

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Properties and storage information

Gene name
HNRNPUL1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

The E1B-AP5 protein also known as hnRNPUL1 serves mechanical functions by binding to RNA and DNA and it acts as a regulator in transcription and mRNA processing. This protein weighs approximately 88 kDa. Researchers have identified its presence in the nucleus of different cell types reflecting its central role in nucleic acid metabolism. The protein can also intervene in alternative splicing events and influence mRNA stability making it a significant player within the cell's genetic expression regulation matrix.
Biological function summary

E1B-AP5 integrates into cellular processes as a part of the heterogeneous nuclear ribonucleoproteins complex which facilitates various RNA processing mechanisms. It plays an active role in chromatin structure modulation and RNA transport fostering proper gene expression and maintaining cellular homeostasis. Besides supporting the transcription machinery E1B-AP5 engages in RNA export and surveillance where its presence determines the fate of newly synthesized RNAs.

Pathways

Studies have shown that E1B-AP5 is involved in pathways such as the mRNA surveillance and splicing pathways. It collaborates with proteins like HNRNPA1 and HNRNPC in orchestrating the correct assembly and exportation of RNA transcripts. By participating in these critical pathways E1B-AP5 ensures the fidelity of gene expression and supports the cellular stress response mechanisms contributing to proper cellular function and adaptability.

Abnormalities in E1B-AP5 expression correlate with oncogenic transformations and neurodegenerative conditions. In cancer for example dysregulation of E1B-AP5 affects RNA processing and can lead to tumor development with its interaction noted alongside the protein p53 in the context of cellular proliferation control. Understanding E1B-AP5's role in these diseases might provide insights into therapeutic targets particularly where RNA processing and tumor suppressor pathways are implicated.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Target data

See full target information HNRNPUL1
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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