Double Strand Breaks (DSB) Repair Antibody Sampler Kit
cat.: HAK21108
Product Type: Antibody Sampler Kit
Species reactivity: Human
Applications: WB
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Uniprot #: SwissProt: Q13315 Human | Q13315 Human | P78527 Human | P97313 Mouse | P13010 Human | P49959 Human | Q61216 Mouse | Q9JIM0 Rat | O60934 Human | Q9R207 Mouse | Q92878 Human | P70388 Mouse | Q9JIL8 Rat | Q9H9Q4 Human
Entrez Gene: 300711 Rat | 360748 Rat
Images
HAK21108_1.jpg Fig1: Western blot analysis of Phospho-ATM (S1981) on different lysates with Rabbit anti-Phospho-ATM (S1981) antibody (ET1705-50) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 1μM Camptothecin for 1 hour cell lysate
Lane 3: HeLa treated with 1μM Camptothecin for 1 hour cell lysate, then the membrane treated with λpp for 1 hour

Lysates/proteins at 20 µg/Lane.

Predicted band size: 351 kDa
Observed band size: 351 kDa

Exposure time: 3 minutes; ECL: K1801;
3-8% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1705-50) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21108_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells treated with or without 1μM Camptothecin for 1 hour labeling Phospho-ATM (S1981) with Rabbit anti-Phospho-ATM (S1981) antibody (ET1705-50) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-ATM (S1981) antibody (ET1705-50) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HAK21108_3.jpg Fig3: Western blot analysis of Phospho-ATM (S1981) on different lysates with Rabbit anti-Phospho-ATM (S1981) antibody (ET1705-50) at 1/2,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 20μM Etoposide for 2 hours cell lysate
Lane 3: HepG2 cell lysate
Lane 4: HEK-293 cell lysate
Lane 5: MDA-MB-231 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 351 kDa
Observed band size: 351 kDa

Exposure time: 1 minute 2 seconds; ECL: K1801;
3-8% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1705-50) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21108_4.jpg Fig4: Western blot analysis of ATM on different lysates with Rabbit anti-ATM antibody (ET1606-20) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: A549 cell lysate
Lane 3: NCCIT cell lysate
Lane 4: HEK-293 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 351 kDa
Observed band size: 351 kDa

Exposure time: 24 seconds; ECL: K1801;
3-8% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1606-20) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21108_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-ATM antibody (ET1606-20) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-20) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HAK21108_6.jpg Fig6: Western blot analysis of ATM on different lysates with Rabbit anti-ATM antibody (ET1606-20) at 1/1,000 dilution.

Lane 1: A549 cell lysate
Lane 2: CRC cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 351 kDa
Observed band size: 351 kDa

Exposure time: 3 minutes;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1606-20) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21108_7.jpg Fig7: Western blot analysis of DNA PKcs on different lysates with Rabbit anti-DNA PKcs antibody (ET1610-12) at 1/5,000 dilution.

Lane 1: K-562 cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: PC-12 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 469 kDa
Observed band size: 469 kDa

Exposure time: 5 seconds; ECL: K1801;

3-8% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-12) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21108_8.jpg Fig8: Western blot analysis of Ku80 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1610-40, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: MCF-7 cell lysate
Lane 2: A549 cell lysate
HAK21108_9.jpg Fig9: Western blot analysis of Mre11 on different lysates with Rabbit anti-Mre11 antibody (ET1703-99) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: 293T cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 81 kDa
Observed band size: 81 kDa

Exposure time: 25 seconds; ECL: K1801;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1703-99) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21108_10.jpg Fig10: Western blot analysis of Phospho-p95/NBS1 (S343) on different lysates with Rabbit anti-Phospho-p95/NBS1 (S343) antibody (ET1607-5) at 1/1,000 dilution.

Lane 1: HeLa whole cell lysate
Lane 2: HeLa treated with 20μM Etoposide for 2 hours whole cell lysate
Lane 3: Jurkat whole cell lysate
Lane 4: Jurkattreated with 25μM Etoposide for 5 hours whole cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 85 kDa
Observed band size: 95 kDa

Exposure time: 5 minutes;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1607-5) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.
HAK21108_11.jpg Fig11: Western blot analysis of Rad50 on different lysates with Rabbit anti-Rad50 antibody (HA721492) at 1/1,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: K-562 cell lysate (20 µg/Lane)
Lane 3: U-87 MG cell lysate (20 µg/Lane)
Lane 4: U-937 cell lysate (20 µg/Lane)
Lane 5: Caco-2 cell lysate (20 µg/Lane)
Lane 6: Mouse testis tissue lysate (40 µg/Lane)
Lane 7: Rat testis tissue lysate (40 µg/Lane)

Predicted band size: 154 kDa
Observed band size: 154 kDa

Exposure time: 3 minutes;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721492) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
HAK21108_12.jpg Fig12: Western blot analysis of XLF on different lysates with Rabbit anti-XLF antibody (HA721572) at 1/1,000 dilution.

Lane 1: HepG2 cell lysate (20 µg/Lane)
Lane 2: 293T cell lysate (20 µg/Lane)
Lane 3: NCCIT cell lysate (30 µg/Lane)
Lane 4: HeLa cell lysate (30 µg/Lane)
Lane 5: Jurkat cell lysate (30 µg/Lane)

Predicted band size: 33 kDa
Observed band size: 37 kDa

Exposure time: 2 minutes 10 seconds;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721572) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.