Coronavirus Host Cell Attachment and Entry Antibody Sampler Kit

Western blot analysis of ACE2 on different lysates with Rabbit anti-ACE2 antibody (ET1611-58) at 1/1,000 dilution.

Lane 1: HepG2 cell lysate
Lane 2: 293T cell lysate
Lane 3: Rat brain tissue lysate
Lane 4: Rat kidney tissue lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 92 kDa
Observed band size: 100 kDa

Exposure time: 2 minutes; ECL: K1802;

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 (ET1611-58) 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.

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-ACE2 antibody (ET1611-58) 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 (ET1611-58) 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.

Western blot analysis of CD26 on different lysates with Rabbit anti-CD26 antibody (ET1703-93) at 1/5,000 dilution.

Lane 1: LoVo cell lysate
Lane 2: Caco-2 cell lysate
Lane 3: Mouse thymus tissue lysate
Lane 4: Rat thymus tissue lysate
Lane 5: Mouse liver tissue lysate
Lane 6: Rat liver tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 88 kDa
Observed band size: 100/110 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 (ET1703-93) 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.

Western blot analysis of CD13 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 (ET1610-59, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Human heart tissue lysate
Lane 2: U937 cell lysate

Western blot analysis of CD147 on different lysates with Rabbit anti-CD147 antibody (ET1702-58) at 1/5,000 dilution.

Lane 1: HepG2 cell lysate (15 µg/Lane)
Lane 2: NCI-H226 cell lysate (15 µg/Lane)
Lane 3: 786-0 cell lysate (15 µg/Lane)
Lane 4: RAW264.7 cell lysate (15 µg/Lane)
Lane 5: Mouse liver tissue lysate (20 µg/Lane)
Lane 6: Rat liver tissue lysate (20 µg/Lane)

Predicted band size: 42 kDa
Observed band size: 45-60 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 (ET1702-58) 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.

Western blot analysis of EEA1 on different lysates with Rabbit anti-EEA1 antibody (HA722147) at 1/1,000 dilution.

Lane 1: HeLa cell lysate, 20 µg/Lane
Lane 2: Jurkat cell lysate, 20 µg/Lane
Lane 3: JAR cell lysate, 20 µg/Lane
Lane 4: NIH/3T3 cell lysate, 20 µg/Lane
Lane 5: C6 cell lysate, 20 µg/Lane
Lane 6: COS-1 cell lysate, 20 µg/Lane

Predicted band size: 162 kDa
Observed band size: 162 kDa

Exposure time: 12 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 (HA722147) 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.

Immunocytochemistry analysis of JAR cells labeling EEA1 with Rabbit anti-EEA1 antibody (HA722147) 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-EEA1 antibody (HA722147) 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.

Western blot analysis of IFITM1 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 (ER1902-77, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: K562 cell lysate
Lane 2: SHG-44 cell lysate

Western blot analysis of Cathepsin B on different lysates with Rabbit anti-Cathepsin B antibody (ET1704-03) at 1/1,000 dilution.

Lane 1: HepG2 cell lysate
Lane 2: SW480 cell lysate
Lane 3: Daudi cell lysate (negative)

Lysates/proteins at 20 µg/Lane.

Predicted band size: 38 kDa
Observed band size: 43 kDa

Exposure time: 1 minutes 59 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 (ET1704-03) 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.

Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-Cathepsin B 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 (ET1704-03, 1/50) 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.

Western blot analysis of Fragilis on different lysates with Rabbit anti-Fragilis antibody (ET1706-09) at 1/2,000 dilution.

Lane 1: HepG2 cell lysate
Lane 2: HeLa cell lysate
Lane 3: THP-1 cell lysate
Lane 4: 293T cell lysate (negative)

Lysates/proteins at 15 µg/Lane.

Predicted band size: 15 kDa
Observed band size: 15 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 (ET1706-09) 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.

Immunocytochemistry analysis of HeLa cells labeling Fragilis with Rabbit anti-Fragilis antibody (ET1706-09) 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-Fragilis antibody (ET1706-09) 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.

Western blot analysis of VCP 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 (ET1703-56, 1/500) 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: SH-SY5Y cell lysate
Lane 2: HepG2 cell lysate
Lane 3: Hela cell lysate