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AB266827

Human PBK (SPK) knockout HEK-293T cell line

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PBK 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: 1 bp deletion in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
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Western blot - Human PBK (SPK) knockout HEK-293T cell line (AB266827)
  • WB

Lab

Western blot - Human PBK (SPK) knockout HEK-293T cell line (AB266827)

Lanes 1- 2 : Merged signal (red and green). Green - ab75987 observed at 40 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab75987 was shown to react with PBK/SPK in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266827 (knockout cell lysate ab257575) was used. Wild-type HEK-293T and PBK knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab75987 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4° at a 1 in 100000 dilution and a 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-PBK/SPK antibody [EP2520Y] (<a href='/ko/products/primary-antibodies/pbk-spk-antibody-ep2520y-ab75987'>ab75987</a>) at 1/100000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

PBK knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human PBK (SPK) knockout HEK-293T cell line (ab266827)

Predicted band size: 27 kDa,36 kDa

Observed band size: 40 kDa

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Western blot - Human PBK (SPK) knockout HEK-293T cell line (AB266827)
  • WB

Lab

Western blot - Human PBK (SPK) knockout HEK-293T cell line (AB266827)

Lanes 1- 2 : Merged signal (red and green). Green - ab236871 observed at 40 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab236871 was shown to react with PBK/SPK in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266827 (knockout cell lysate ab257575) was used. Wild-type HEK-293T and PBK knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab236871 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4° at a 1 in 1000 dilution and a 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-PBK/SPK antibody [EPR21982] (<a href='/ko/products/primary-antibodies/pbk-spk-antibody-epr21982-ab236871'>ab236871</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

PBK knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human PBK (SPK) knockout HEK-293T cell line (ab266827)

Predicted band size: 36 kDa

Observed band size: 40 kDa

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Cell Culture - Human PBK (SPK) knockout HEK-293T cell line (AB266827)
  • Cell Culture

Unknown

Cell Culture - Human PBK (SPK) knockout HEK-293T cell line (AB266827)

Representative images of PBK 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 Sequencing - Human PBK (SPK) knockout HEK-293T cell line (AB266827)
  • Sanger seq

Unknown

Sanger Sequencing - Human PBK (SPK) knockout HEK-293T cell line (AB266827)

Homozygous : 1 bp deletion in exon 2

주요 정보

세포 유형

HEK-293T

Species or organism

Human

조직

Kidney

제형

Liquid

form

Knockout 검증

Sanger Sequencing,Western blot

Mutation 정보

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 2

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보완 제품

Primary Antibodies

AB32529

Anti-S6K1 antibody [E343]

KO Validated|RabMAb|Recombinant|Lab Essentials|20ul selling size

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S6K1 antibody [E343] (AB32529)

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  • 1
Primary Antibodies

AB178440

Anti-Ref2P antibody

Western blot - Anti-Ref2P antibody (AB178440)

  • 4
  • 5
Primary Antibodies

AB177472

Anti-ULK1 antibody [EPR6153(2)]

20ul selling size|RabMAb|Recombinant

Western blot - Anti-ULK1 antibody [EPR6153(2)] (AB177472)

  • 0
  • 0
Primary Antibodies

AB303488

Anti-ATG12 antibody [EPR27485-89]

BOND RX™ Validated|20ul selling size|RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG12 antibody [EPR27485-89] (AB303488)

  • 0
  • 0
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Reactivity 정보

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

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.

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제품 구성

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특성 및 보관 정보

유전자명
PBK
유전자 편집 종류
Knockout
유전자 편집 방식
CRISPR technology
Knockout 검증
Sanger Sequencing, Western blot
Zygosity
Homozygous
배송 시 보관 조건
Dry Ice
적절한 단기 보관 조건
-196°C
적절한 장기 보관 조건
-196°C
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추가 정보

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

PBK/SPK also known as PDZ-binding kinase or Cdc2-related kinase is a serine/threonine protein kinase with an approximate molecular weight of 47 kDa. This protein is involved in cell division playing an important role in mitotic processes. PBK/SPK is expressed in various tissues including significant levels in testis and placenta often in proliferating cell types. It also exhibits higher expression in cancerous tissues suggesting a role in tumorigenesis.
Biological function summary

PBK/SPK facilitates cell cycle progression and cellular proliferation. This protein is not typically part of larger complexes but it partners with other molecules during cell division to ensure proper mitotic spindle formation and chromosome segregation. Its expression levels increase during the G2/M phase of the cell cycle indicating its involvement in cell cycle regulation. In cancer it often shows overexpression contributing to unregulated proliferation of cancer cells.

Pathways

PBK/SPK is associated with critical cell division pathways such as the MAPK signaling pathway. This involvement impacts cellular responses to growth signals and stress. PBK can phosphorylate and interact with proteins like p38 MAPK which further modulates signal transduction related to cellular growth and survival. Through these pathways PBK/SPK helps manage the balance between cell division and apoptosis particularly under stress conditions.

PBK/SPK has strong correlations with cancer especially glioblastoma and cervical cancer. Overexpression of PBK often links to poor prognosis in these conditions. In cancerous settings PBK interacts with proteins such as p53 and c-Myc influencing tumor progression and resistance to therapy. PBK can modulate oncogenic pathways providing a potential target for therapeutic intervention in tumor proliferation and survival.
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품질 관리

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

초기 취급 지침

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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>

Subculture 가이드라인
  • 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.
배양 배지

DMEM (High Glucose) + 10% FBS

동결보존 배지

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

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제품 프로토콜

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Product promise

저희는 고품질 시약으로 고객님의 연구를 서포트하고자 최선을 다하고 있으며, 모든 단계에서 함께 하겠습니다. 만약 제품이 기대한 성능을 보이지 않을 경우에도 Product Promise로 보호받으실 수 있습니다.
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