HAK21164

Cuproptosis Essentials Antibody Sampler Kit

cat.: HAK21164

Product Type:	Antibody Sampler Kit
Species reactivity:	Human, Mouse
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.
Concentration:	1ug/ul
Uniprot #:	SwissProt: P10109 Human \| P46656 Mouse \| P24483 Rat \| P10515 Human \| Q8BMF4 Mouse \| P08461 Rat \| O43766 Human \| Q99M04 Mouse \| Q5XIH4 Rat \| O15431 Human \| Q8K211 Mouse \| Q9JK41 Rat \| P09622 Human \| O08749 Mouse

Images

Fig1: Western blot analysis of LIAS on different lysates with Rabbit anti-LIAS antibody (HA722085) at 1/2,000 dilution.

Lane 1: K-562 cell lysate (20 µg/Lane)
Lane 2: LO2 cell lysate (20 µg/Lane)
Lane 3: Raji cell lysate (20 µg/Lane)
Lane 4: MCF7 cell lysate (20 µg/Lane)
Lane 5: Jurkat cell lysate (20 µg/Lane)
Lane 6: HepG2 cell lysate (20 µg/Lane)
Lane 7: PC-3M cell lysate (20 µg/Lane)
Lane 8: C2C12 cell lysate (20 µg/Lane)
Lane 9: L6 cell lysate (20 µg/Lane)
Lane 10: Mouse liver tissue lysate (40 µg/Lane)
Lane 11: Mouse colon tissue lysate (40 µg/Lane)
Lane 12: Mouse brain tissue lysate (40 µg/Lane)
Lane 13: Rat liver tissue lysate (40 µg/Lane)
Lane 14: Rat testis tissue lysate (40 µg/Lane)

Predicted band size: 42 kDa
Observed band size: 39 kDa

Exposure time: 42 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 (HA722085) 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.

Fig2: Western blot analysis of Pyruvate Dehydrogenase E2 on different lysates with Rabbit anti-Pyruvate Dehydrogenase E2 antibody (HA721267) at 1/1,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-Pyruvate Dehydrogenase E2 KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 69 kDa
Observed band size: 69 kDa

Exposure time: 4 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 (HA721267) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig3: Western blot analysis of SLC31A1 / CTR1 on different lysates with Rabbit anti-SLC31A1 / CTR1 antibody (HA720005) at 1/1,000 dilution.

Lane 1: A549 cell lysate (20 µg/Lane)
Lane 2: Neuro-2a cell lysate (20 µg/Lane)
Lane 3: C6 cell lysate (20 µg/Lane)
Lane 4: Mouse liver tissue lysate (40 µg/Lane)
Lane 5: Rat brain tissue lysate (40 µg/Lane)

Predicted band size: 21 kDa
Observed band size: 30 kDa

Exposure time: 6 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 (HA720005) 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.

Fig4: Western blot analysis of Lipoamide Dehydrogenase on different lysates with Rabbit anti-Lipoamide Dehydrogenase antibody (HA721512) at 1/1,000 dilution.

Lane 1: Jurkat cell lysate (20 µg/Lane)
Lane 2: HeLa cell lysate (20 µg/Lane)
Lane 3: 293T cell lysate (20 µg/Lane)
Lane 4: MCF7 cell lysate (20 µg/Lane)
Lane 5: K-562 cell lysate (20 µg/Lane)
Lane 6: SK-Br-3 cell lysate (20 µg/Lane)
Lane 7: U-937 cell lysate (20 µg/Lane)
Lane 8: HepG2 cell lysate (20 µg/Lane)
Lane 9: NIH/3T3 cell lysate (20 µg/Lane)
Lane 10: Mouse liver tissue lysate (40 µg/Lane)

Predicted band size: 54 kDa
Observed band size: 54 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 (HA721512) 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.

Fig5: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Lipoamide Dehydrogenase antibody (HA721512) at 1/5,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (HA721512) at 1/5,000 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.

Fig6: Immunocytochemistry analysis of HeLa cells labeling Lipoamide Dehydrogenase with Rabbit anti-Lipoamide Dehydrogenase antibody (HA721512) at 1/200 dilution.

Cells were fixed in 100% precooled methanol 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-Lipoamide Dehydrogenase antibody (HA721512) at 1/200 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.

Fig7: Western blot analysis of ADX on different lysates with Rabbit anti-ADX antibody (HA721329) at 1/500 dilution.

Lane 1: SK-Br-3 cell lysate (15 µg/Lane)
Lane 2: K-562 cell lysate (15 µg/Lane)
Lane 3: HepG2 cell lysate (15 µg/Lane)
Lane 4: 293T cell lysate (15 µg/Lane)

Predicted band size: 19 kDa
Observed band size: 13 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 (HA721329) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

Fig8: Immunocytochemistry analysis of C2C12 cells labeling LIAS with Rabbit anti-LIAS antibody (HA722085) at 1/100 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-LIAS antibody (HA722085) at 1/100 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.

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