Human KDM5C (Jarid1C / SMCX) knockout HEK-293T cell line
- Advanced Validation
- 각 태그의 뜻
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KDM5C 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 deletion in exon 2 and 1 bp insertion in exon 2 and Insertion of the selection cassette 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'.
대체 명칭 보기
KDM5C, Histone demethylase JARID1C, [histone H3]-trimethyl-L-lysine(4) demethylase 5C, JARID1C, Lysine-specific demethylase 5C, Protein Xe169, DXS1272E, Jumonji/ARID domain-containing protein 1C, SMCX, Protein SmcX, XE169
- WB
Lab
Western blot - Human KDM5C (Jarid1C / SMCX) knockout HEK-293T cell line (AB266251)
Blocking and Diluting buffer and concentration : 5% NFDM/TBST
Lanes 1-4 : Merged signal (red and green). Green - ab259913 observed at 180 kDa. Red - loading control ab8245 (Mouse monoclonal [6C5] to GAPDH) observed at 36 kDa.
ab259913 Anti-KDM5C / Jarid1C / SMCX antibody [EPR23932-18] was shown to specifically react with KDM5C / Jarid1C / SMCX in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266251 (knockout cell lysate ab257494) and ab266252 (knockout cell lysate ab257495) were used. Wild-type and KDM5C / Jarid1C / SMCX knockout samples were subjected to SDS-PAGE. ab259913 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4℃ overnight 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 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-KDM5C / Jarid1C / SMCX antibody [EPR23932-18] (<a href='/ko/products/primary-antibodies/kdm5c-jarid1c-smcx-antibody-epr23932-18-ab259913'>ab259913</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg
Lane 2:
Western blot - Human KDM5C (Jarid1C / SMCX) knockout HEK-293T cell line (<a href='/ko/products/cell-lines/human-kdm5c-jarid1c-smcx-knockout-hek-293t-cell-line-ab266252'>ab266252</a>) at 20 µg
Lane 3:
Western blot - Human KDM5C (Jarid1C / SMCX) knockout HEK-293T cell line (ab266251) at 20 µg
Lane 4:
HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Secondary
All lanes:
Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/ko/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>)
Predicted band size: 175 kDa
Observed band size: 180 kDa
false
- WB
Lab
Western blot - Human KDM5C (Jarid1C / SMCX) knockout HEK-293T cell line (AB266251)
Lanes 1 - 2 : Merged signal (red and green). Green - ab34718 observed at 175 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab34718 was shown to react with KDM5C / Jarid1C / SMCX in wild-type HEK-293T cells in western blot with loss of signal observed in KDM5C knockout cell line ab266251 (KDM5C knockout cell lysate ab257494). Wild-type and KDM5C knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab34718 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4° at a 1 in 250 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-KDM5C / Jarid1C / SMCX antibody (<a href='/ko/products/primary-antibodies/kdm5c-jarid1c-smcx-antibody-ab34718'>ab34718</a>) at 1/250 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
KDM5C knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human KDM5C (Jarid1C / SMCX) knockout HEK-293T cell line (ab266251)
Predicted band size: 175 kDa
Observed band size: 175 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human KDM5C (Jarid1C / SMCX) knockout HEK-293T cell line (AB266251)
Allele-1 : 1 bp deletion in exon 2
- Sanger seq
Unknown
Sanger Sequencing - Human KDM5C (Jarid1C / SMCX) knockout HEK-293T cell line (AB266251)
Allele-3 : Insertion of the selection cassette in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human KDM5C (Jarid1C / SMCX) knockout HEK-293T cell line (AB266251)
Allele-2 : 1 bp insertion in exon 2.
- WB
Supplier Data
Western blot - Human KDM5C (Jarid1C / SMCX) knockout HEK-293T cell line (AB266251)
This data was developed using ab259913, the same antibody clone in a different buffer formulation.
Blocking and Diluting buffer and concentration : 5% NFDM/TBST
Lanes 1-4 : Merged signal (red and green). Green - ab259913 observed at 180 kDa. Red - loading control ab8245 (Mouse monoclonal [6C5] to GAPDH) observed at 36 kDa.
Lanes 1-3 : ab259913 Anti-KDM5C / Jarid1C / SMCX antibody [EPR23932-18] was shown to specifically react with KDM5C / Jarid1C / SMCX in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266251 (knockout cell lysate ab257494) and ab266252 (knockout cell lysate ab257495) were used. Wild-type and KDM5C / Jarid1C / SMCX knockout samples were subjected to SDS-PAGE. ab259913 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4℃ overnight 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 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-KDM5C / Jarid1C / SMCX antibody [EPR23932-18] (<a href='/ko/products/primary-antibodies/kdm5c-jarid1c-smcx-antibody-epr23932-18-ab259913'>ab259913</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg
Lane 2:
Western blot - Human KDM5C (Jarid1C / SMCX) knockout HEK-293T cell line (<a href='/ko/products/cell-lines/human-kdm5c-jarid1c-smcx-knockout-hek-293t-cell-line-ab266252'>ab266252</a>) at 20 µg
Lane 3:
Western blot - Human KDM5C (Jarid1C / SMCX) knockout HEK-293T cell line (ab266251) at 20 µg
Lane 4:
HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Secondary
All lanes:
Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/ko/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution
Predicted band size: 175 kDa
Observed band size: 180 kDa
false
보완 제품
Cell Lines & Lysates
AB266900
Human TUBB3 (beta III Tubulin) knockout HCT116 cell line
Advanced Validation
- 0
- 0
Reactivity 정보
제품 세부 정보
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.
제품 구성
특성 및 보관 정보
유전자명
유전자 편집 종류
유전자 편집 방식
Knockout 검증
배송 시 보관 조건
적절한 단기 보관 조건
적절한 장기 보관 조건
품질 관리
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
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.
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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