Anti-Ly-6A/E (Sca-1) antibody [EPR22502-38]

• Flow Cyt

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Flow Cytometry - Anti-Ly-6A/E (Sca-1) antibody [EPR22502-38] (AB219794)

Flow cytometric analysis of mouse bone marrow cells labeling Ly-6A/E (Sca1) with ab219794 at 1/500 (right panel) compared with a Rabbit monoclonal IgG (ab172730) (left panel). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

• Flow Cyt

Lab

Flow Cytometry - Anti-Ly-6A/E (Sca-1) antibody [EPR22502-38] (AB219794)

Flow cytometry staining of C57 BL/6 mouse splenocytes with ab219794 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left). Cells were incubated for 30 mins on ice in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab219794 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (1x 10⁶ in 100 µl at 0.2 μg/ml (1/2500 dilution)) for 30 mins on ice. The cells were simultaneously stained with C-Kit.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was incubated at 1/4000 dilution for 30 mins on ice

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on live cells.

• WB

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Western blot - Anti-Ly-6A/E (Sca-1) antibody [EPR22502-38] (AB219794)

This blot was developed using a higher sensitivity ECL substrate.

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Ly-6A/E (Sca-1) antibody [EPR22502-38] (ab219794) at 1/1000 dilution

All lanes:

C2C12 (mouse myoblasts myoblast), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 14 kDa

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• Flow Cyt

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Flow Cytometry - Anti-Ly-6A/E (Sca-1) antibody [EPR22502-38] (AB219794)

Flow cytometric analysis of mouse splenocytes treated with 2.5μg/ml Concanavalin A (ConA) for 72h labeling Ly-6A/E (Sca-1) with ab219794 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

• WB

Unknown

Western blot - Anti-Ly-6A/E (Sca-1) antibody [EPR22502-38] (AB219794)

Negative control : EL4 (PMID : 2430016).

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Ly-6A/E (Sca-1) antibody [EPR22502-38] (ab219794) at 1/1000 dilution

Lane 1:

Mouse kidney tissue lysate at 20 µg

Lane 2:

EL4 (mouse lymphoma T lymphocyte), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 14 kDa

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Exposure time: 48s