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AB265868

Human HPDL knockout HeLa cell line

Human HPDL knockout HeLa cell line

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HPDL KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 2 bp deletion in exon 1. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.

別名を表示する

4 HPPD L, 4-hydroxyphenylpyruvate dioxygenase-like, 4-hydroxyphenylpyruvate dioxygenase-like protein, Gloxd1, Glyoxalase domain containing 1, Glyoxalase domain-containing protein 1, HPDL_HUMAN

3 Images
Western blot - Human HPDL knockout HeLa cell line (AB265868)
  • WB

Lab

Western blot - Human HPDL knockout HeLa cell line (AB265868)

Lanes 1-3 : Merged signal (red and green). Green - ab174841 observed at 39 kDa. Red - loading control ab7291 observed at 50 kDa.

ab174841 Anti-HPDL antibody [EPR11691] - C-terminal was shown to specifically react with HPDL in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265868 (knockout cell lysate ab257995) was used. Wild-type and HPDL knockout samples were subjected to SDS-PAGE. ab174841 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-HPDL antibody [EPR11691] - C-terminal (<a href='/products/primary-antibodies/hpdl-antibody-epr11691-c-terminal-ab174841'>ab174841</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

HPDL knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human HPDL knockout HeLa cell line (ab265868)

Lane 3:

Caco-2 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 39 kDa

Observed band size: 39 kDa

false

Sanger Sequencing - Human HPDL knockout HeLa cell line (AB265868)
  • Sanger seq

Unknown

Sanger Sequencing - Human HPDL knockout HeLa cell line (AB265868)

Allele-1 : 2 bp deletion in exon 1.

Sanger Sequencing - Human HPDL knockout HeLa cell line (AB265868)
  • Sanger seq

Unknown

Sanger Sequencing - Human HPDL knockout HeLa cell line (AB265868)

Allele-2 : 1 bp insertion in exon 1.

Key facts

細胞タイプ

HeLa

生物種

Human

組織

Cervix

製品の状態

Liquid

form

ノックアウト検証方法

Sanger Sequencing,Western blot

ノックアウト変異

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 2 bp deletion in exon 1

疾病

Adenocarcinoma

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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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製品の詳細

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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製品内容

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出荷温度及び保存条件

遺伝子名
HPDL
遺伝子編集のタイプ
Knockout
遺伝子編集の方法
CRISPR technology
ノックアウト検証方法
Sanger Sequencing, Western blot
出荷温度
Dry Ice
短期保存温度
-196°C
長期保存温度
-196°C
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取り扱い方法

初回取り扱いガイドライン

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.

継代培養ガイドライン
  • 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.
培養培地

DMEM (High Glucose) + 10% FBS

凍結保存培地

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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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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製品プロトコール

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ターゲットの情報

See full target information HPDL
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組合せ製品

Primary Antibodies

AB174841

Anti-HPDL antibody [EPR11691] - C-terminal

RabMAb|Recombinant|KO Validated

Immunoprecipitation - Anti-HPDL antibody [EPR11691] - C-terminal (AB174841)

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Proteins & Peptides

AB195163

Recombinant Human ZMYND8 protein

SDS-PAGE - Recombinant Human ZMYND8 protein (AB195163)

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  • 0
Proteins & Peptides

AB313361

Recombinant Human WFDC2 Protein (His tag)

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

  • 0
  • 0
Proteins & Peptides

AB217407

Recombinant human Vitronectin/S-Protein (Animal Free)

  • 0
  • 0

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