Glutathione metabolism Antibody Sampler Kit
cat.: HAK21181
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.
Concentration: 1ug/ul
Uniprot #: SwissProt: P07203 Human | P00390 Human | P47791 Mouse | P70619 Rat | Q9UPY5 Human | Q9WTR6 Mouse | P48507 Human | O09172 Mouse | P48508 Rat | P48506 Human | P19468 Rat | P09211 Human | P19157 Mouse | P04906 Rat | Q9Y2Q3 Human | P10620 Human | Q91VS7 Mouse | P08011 Rat | P33527 Human
Entrez Gene: 310392 Rat
Images
HAK21181_1.jpg Fig1: Western blot analysis of Glutathione peroxidase 1 on different lysates with Mouse anti-Glutathione peroxidase 1 antibody (HA601200) at 1/1,000 dilution.

Lane 1: HEK-293 cell lysate (20 µg/Lane)
Lane 2: HepG2 cell lysate (20 µg/Lane)
Lane 3: SH-SY5Y cell lysate (20 µg/Lane)
Lane 4: HT-29 cell lysate (20 µg/Lane)
Lane 5: Human liver tissue lysate (40 µg/Lane)
Lane 6: Human kidney tissue lysate (40 µg/Lane)

Predicted band size: 22 kDa
Observed band size: 22 kDa

Exposure time: 24 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 (HA601200) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21181_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human liver tissue with Mouse anti-Glutathione peroxidase 1 antibody (HA601200) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA601200) 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.
HAK21181_3.jpg Fig3: Western blot analysis of Glutathione Reductase on different lysates with Rabbit anti-Glutathione Reductase antibody (HA721989) at 1/2,000 dilution.

Lane 1: HepG2 cell lysate (20 µg/Lane)
Lane 2: A549 cell lysate (20 µg/Lane)
Lane 3: K-562 cell lysate (20 µg/Lane)
Lane 4: Jurkat cell lysate (20 µg/Lane)
Lane 5: Mouse liver tissue lysate (40 µg/Lane)
Lane 6: Mouse brain tissue lysate (40 µg/Lane)
Lane 7: Mouse kidney tissue lysate (40 µg/Lane)
Lane 8: Rat lung tissue lysate (40 µg/Lane)
Lane 9: Rat kidney tissue lysate (40 µg/Lane)

Predicted band size: 56 kDa
Observed band size: 54 kDa

Exposure time: 10 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 (HA721989) 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.
HAK21181_4.jpg Fig4: Western blot analysis of Glutathione Reductase on different lysates with Rabbit anti-Glutathione Reductase antibody (HA721989) at 1/2,000 dilution.

Lane 1: HCT 116-si NT cell lysate
Lane 2: HCT 116-si Glutathione Reductase cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 56 kDa
Observed band size: 54 kDa

Exposure time: 20 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 (HA721989) 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.
HAK21181_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-xCT / SLC7A11 antibody (HA724114) at 1/50 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA724114) at 1/50 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.
HAK21181_6.jpg Fig6: Immunocytochemistry analysis of A549 cells labeling xCT / SLC7A11 with Rabbit anti-xCT / SLC7A11 antibody (HA724114) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-xCT / SLC7A11 antibody (HA724114) 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 (HA601187 , 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.
HAK21181_7.jpg Fig7: Western blot analysis of GCLM on different lysates with Rabbit anti-GCLM antibody (ET1705-87) at 1/1,000 dilution.

Lane 1: A431 cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: PC-12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 31 kDa
Observed band size: 31 kDa

Exposure time: 59 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 (ET1705-87) 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.
HAK21181_8.jpg Fig8: Immunofluorescence analysis of frozen mouse hippocampus tissue labeling GCLM with Rabbit anti-GCLM antibody (ET1705-87).

The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET1705-87, green) at 1/50 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.
HAK21181_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-GCLM antibody (ET1705-87) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1705-87) at 1/200 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.
HAK21181_10.jpg Fig10: Western blot analysis of GCLC on different lysates with Rabbit anti-GCLC antibody (ET1704-38) at 1/5,000 dilution.

Lane 1: A549-si NT cell lysate
Lane 2: A549-si GCLC cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 73 kDa
Observed band size: 73 kDa

Exposure time: 2 minutes; 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 (ET1704-38) 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.
HAK21181_11.jpg Fig11: Western blot analysis of MRP1 on different lysates with Rabbit anti-MRP1 antibody (ET1704-56) at 1/1,000 dilution.

Lane 1: A549 cell lysate
Lane 2: A549 cell lysate (no heat)
Lane 3: HepG2 cell lysate
Lane 4: HepG2 cell lysate (no heat)

Notice: no heat means the lysate is not boiled.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 172 kDa
Observed band size: 220 kDa

Exposure time: 3 minutes; 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 (ET1704-56) 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.
HAK21181_12.jpg Fig12: Western blot analysis of MRP1 on different lysates with Rabbit anti-MRP1 antibody (ET1704-56) at 1/1,000 dilution.

Lane 1: A549-si NT cell lysate (no heat)
Lane 2: A549-si MRP1 cell lysate (no heat)

Notice: no heat means the lysate is not boiled.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 172 kDa
Observed band size: 220 kDa

Exposure time: 3 minutes; 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 (ET1704-56) 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.
HAK21181_13.jpg Fig13: Immunocytochemistry analysis of A549 cells labeling MRP1 with Rabbit anti-MRP1 antibody (ET1704-56) 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-MRP1 antibody (ET1704-56) 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”.