Senescence Marker Antibody Sampler Kit
cat.: HAK21082
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: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Uniprot #: SwissProt: P42771 Human | P38936 Human | P16104 Human | P27661 Mouse | P20700 Human | P14733 Mouse | P70615 Rat | P09429 Human | P63158 Mouse | P63159 Rat | P05231 Human | P08505 Mouse | P01375 Human | P06804 Mouse | P08254 Human | P28862 Mouse | P03957 Rat
Unigene: 2850 Rat
Images
HAK21082_1.jpg Fig1: Western blot analysis of Phospho-Histone H2A.X (S139) on different lysates with Rabbit anti-Phospho-Histone H2A.X (S139) antibody (ET1602-2) at 1/2,000 dilution.

Lane 1: HeLa whole cell lysate
Lane 2: HeLa treated with 20μM Etoposide for 2 hours whole cell lysate
Lane 3: HeLa whole cell lysate
Lane 4: HeLa treated with UV for 2 hours whole cell lysate
Lane 5: NIH/3T3 whole cell lysate
Lane 6: NIH/3T3 treated with 25μM Etoposide for 5 hours whole cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 15 kDa
Observed band size: 15/20 kDa

Exposure time: 53 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 (ET1602-2) at 1/2,000 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.
HAK21082_2.jpg Fig2: Western blot analysis of HMGB1 on different lysates with Rabbit anti-HMGB1 antibody (ET1601-2) at 1/50,000 dilution and competitor's antibody at 1/10,000 dilution.

Lane 1: HepG2 cell lysate
Lane 2: HeLa cell lysate
Lane 3: HCT 116 cell lysate
Lane 4: A549 cell lysate
Lane 5: Jurkat cell lysate
Lane 6: C2C12 cell lysate
Lane 7: C6 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 25 kDa
Observed band size: 25 kDa

Exposure time: 21 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 (ET1601-2) at 1/50,000 dilution and competitor's antibody at 1/10,000 dilution were 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.
HAK21082_3.jpg Fig3: Immunocytochemistry analysis of HeLa cells labeling HMGB1 with Rabbit anti-HMGB1 antibody (ET1601-2) at 1/500 dilution and competitor's antibody at 1/250 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-HMGB1 antibody (ET1601-2) at 1/500 dilution and competitor's antibody at 1/250 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.
HAK21082_4.jpg Fig4: Western blot analysis of HMGB1 on different lysates with Rabbit anti-HMGB1 antibody (ET1601-2) at 1/20,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: HCT 116 cell lysate (20 µg/Lane)
Lane 3: A549 cell lysate (20 µg/Lane)
Lane 4: HepG2 cell lysate (20 µg/Lane)
Lane 5: Jurkat cell lysate (20 µg/Lane)
Lane 6: Mouse thymus tissue lysate (20 µg/Lane)
Lane 7: Rat spleen tissue lysate (30 µg/Lane)

Predicted band size: 25 kDa
Observed band size: 25 kDa

Exposure time: 3 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 (ET1601-2) at 1/20,000 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.
HAK21082_5.jpg Fig5: Western blot analysis of TNF alpha on different lysates with Rabbit anti-TNF alpha antibody (HA722022) at 1/1,000 dilution.

Lane 1: THP-1 cell lysate
Lane 2: THP-1 treated with 500ng/mL LPS for 24 hours then add 300ng/mL BFA for 20 hours cell lysate
Lane 3: RAW264.7 cell lysate
Lane 4: RAW264.7 treated with 100ng/mL LPS for 7 hours, add 300ng/mL BFA for 3 hours cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 26 kDa
Observed band size: 26/50/17 kDa

Exposure time: 1 minute;

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 (HA722022) 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.