γ Secretase Antibody Sampler Kit
cat.: HAK21087
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: Q92542 Human | P57716 Mouse | Q9NZ42 Human | Q9CQR7 Mouse | O88777 Rat | P49768 Human | P49769 Mouse | P97887 Rat | P49810 Human | Q61144 Mouse | O88777 Rat
Images
HAK21087_1.jpg Fig1: Western blot analysis of Presenilin 2/AD5 on different lysates with Rabbit anti-Presenilin 2/AD5 antibody (HA722243) at 1/1,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: HepG2 cell lysate (20 µg/Lane)
Lane 3: A549 cell lysate (20 µg/Lane)
Lane 4: 293T cell lysate (20 µg/Lane)
Lane 5: SH-SY5Y cell lysate (20 µg/Lane)
Lane 6: RAW264.7 cell lysate (20 µg/Lane)
Lane 7: Neuro-2a cell lysate (20 µg/Lane)
Lane 8: C2C12 cell lysate (20 µg/Lane)
Lane 9: NIH/3T3 cell lysate (20 µg/Lane)
Lane 10: PC-12 cell lysate (20 µg/Lane)
Lane 11: C6 cell lysate (20 µg/Lane)
Lane 12: Human brain tissue lysate (40 µg/Lane)
Lane 13: Mouse brain tissue lysate (40 µg/Lane)
Lane 14: Rat liver tissue lysate (40 µg/Lane)

Predicted band size: 50 kDa
Observed band size: 23 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 (HA722243) 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.
HAK21087_2.jpg Fig2: Presenilin 2/AD5 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA722243 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722243 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: HA722243 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA722243 in HeLa cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 59 seconds; ECL: K1801
HAK21087_3.jpg Fig3: Western blot analysis of Presenilin 1/PS-1 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 (HA500219, 1/1,000) was used in 5% BSA 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.
Positive control:
Lane 1: NIH/3T3 cell lysate
Lane 2: Raji cell lysate
Lane 3: MCF-7 cell lysate
Lane 4: Daudi cell lysate
HAK21087_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human small intestine tissue using anti-Presenilin 1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA500219, 1/400) for 30 minutes 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.
HAK21087_5.jpg Fig5: Immunofluorescence analysis of frozen rat brain tissue with Rabbit anti-PEN2 antibody (ET7109-26) at 1/100 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET7109-26, green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
HAK21087_6.jpg Fig6: Immunofluorescence analysis of paraffin-embedded mouse testis tissue labeling PEN2 with Rabbit anti-PEN2 antibody (ET7109-26) 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET7109-26, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.