Sequestosome Signaling Antibody Sampler Kit
cat.: HAK21100
| 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. |
| Uniprot #: | SwissProt: Q13501 Human | Q64337 Mouse | O08623 Rat | Q9Y4K3 Human | P70196 Mouse | B5DF45 Rat | P0CG47 Human | P0CG48 Human | P62979 Human | P62987 Human | P0CG49 Mouse | P62991 Mouse | P0CG51 Rat | P62989 Rat | P04629 Human | Q3UFB7 Mouse | P35739 Rat | Q16236 Human | Q60795 Mouse | Q14145 Human | Q9Z2X8 Mouse | P57790 Rat |
Images
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Fig1:
Western blot analysis of SQSTM1 / p62 on different lysates with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/5,000 dilution. Lane 1: SH-SY5Y cell lysate Lane 2: PANC-1 cell lysate Lane 3: HeLa cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 48 kDa Observed band size: 62 kDa Exposure time: 1 minute; 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 (HA721171) at 1/5,000 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. |
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Fig2:
Western blot analysis of SQSTM1 / p62 on different lysates with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/2,000 dilution. Lane 1: A549-si NT cell lysate Lane 2: A549-si SQSTM1 / p62 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 48 kDa Observed band size: 62 kDa Exposure time: 17 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 (HA721171) 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. |
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Fig3:
Immunocytochemistry analysis of C2C12 cells labeling SQSTM1 / p62 with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/1,000 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-SQSTM1 / p62 antibody (HA721171) at 1/1,000 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. |
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Fig4:
Immunocytochemistry analysis of PC-12 cells labeling SQSTM1 / p62 with Rabbit anti-SQSTM1 / p62 antibody (HA721171) 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-SQSTM1 / p62 antibody (HA721171) 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. |
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Fig5:
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-SQSTM1 / p62 antibody (HA721171) 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 (HA721171) 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. |
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Fig6:
Western blot analysis of TRAF6 on different lysates with Rabbit anti-TRAF6 antibody (HA722857) at 1/2,000 dilution. Lane 1: K-562 cell lysate Lane 2: 293T cell lysate Lane 3: HeLa cell lysate Lane 4: COS-1 cell lysate Lane 5: CTLL-2 cell lysate Lane 6: RAW264.7 cell lysate Lane 7: PC-12 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 60 kDa Observed band size: 60 kDa Exposure time: 1 minute; 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 (HA722857) 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. |
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Fig7:
Flow cytometric analysis of RAW264.7 cells labeling TRAF6. Cells were fixed and permeabilized. Then stained with the primary antibody (HA722857, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig8:
Western blot analysis of TrkA on different lysates with Rabbit anti-TrkA antibody (ET1608-44) at 1/5,000 dilution. Lane 1: TF-1 cell lysate Lane 2: 293T cell lysate (negative) Lane 3: Mouse brain tissue lysate Lane 4: Rat brain tissue lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 88 kDa Observed band size: 140 kDa Exposure time: 1 minute 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 (ET1608-44) 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. |
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Fig9:
Western blot analysis of Nrf2 on different lysates with Rabbit anti-Nrf2 antibody (HA721432) at 1/1,000 dilution. Lane 1: HeLa whole cell lysate Lane 2: HeLa treated with 2μM MG-132 for 18 hours whole cell lysate Lane 3: HCT 116 whole cell lysate Lane 4: HCT 116 treated with 25μM MG-132 for 4 hours whole cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 68 kDa Observed band size: 110 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 (HA721432) 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. |
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Fig10:
Western blot analysis of Keap1 on different lysates with Rabbit anti-Keap1 antibody (HA721525) at 1/1,000 dilution. Lane 1: HepG2 cell lysate Lane 2: HeLa cell lysate Lane 3: MCF7 cell lysate Lane 4: A431 cell lysate Lane 5: Jurkat cell lysate Lane 6: HEK-293 cell lysate Lane 7: Daudi cell lysate Lane 8: A549 cell lysate Lane 9: NIH/3T3 cell lysate Lane 10: PC-12 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 70 kDa Observed band size: 55-70 kDa Exposure time: 2 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 (HA721525) 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. |
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Fig11:
Western blot analysis of Keap1 on different lysates with Rabbit anti-Keap1 antibody (HA721525) at 1/1,000 dilution. Lane 1: HepG2-si NT cell lysate Lane 2: HepG2-si Keap1 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 70 kDa Observed band size: 55-70 kDa Exposure time: 1 minute 55 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 (HA721525) at 1/1,000 dilution was used in 5% BSA at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig12:
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Keap1 antibody (HA721525) 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 (HA721525) 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. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.