Anti-ALDH1L1 antibody [3E9] - Astrocyte Marker

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALDH1L1 antibody [3E9] - Astrocyte Marker (AB56777)

ALDH1L1 antibody (ab56777) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human kidney.

This image was generated using the ascites version of the product.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-ALDH1L1 antibody [3E9] - Astrocyte Marker (AB56777)

ICC/IF image of ab56777 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab56777, 1μg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

This image was generated using the ascites version of the product.

• Flow Cyt

Unknown

Flow Cytometry - Anti-ALDH1L1 antibody [3E9] - Astrocyte Marker (AB56777)

Overlay histogram showing HepG2 cells stained with ab56777 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56777, 0.5μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was Mouse IgG3 [MG3-35] (ab18394, 1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

This image was generated using the ascites version of the product.

• IP

Unknown

Immunoprecipitation - Anti-ALDH1L1 antibody [3E9] - Astrocyte Marker (AB56777)

ALDH1L1 was immunoprecipitated using 0.5mg Mouse Liver whole tissue extract, 5μg of Mouse monoclonal to ALDH1L1 and 50μl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Liver whole tissue extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40μl SDS loading buffer and incubated for 10min at 70^oC; 10μl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab56777.
Secondary : Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Band : 99kDa : ALDH1L1; non specific - 60kDa : We are unsure as to the identity of this extra band.

This image was generated using the ascites version of the product.

All lanes:

Immunoprecipitation - Anti-ALDH1L1 antibody [3E9] - Astrocyte Marker (ab56777)

Predicted band size: 99 kDa

false

• WB

Unknown

Western blot - Anti-ALDH1L1 antibody [3E9] - Astrocyte Marker (AB56777)

ALDH1L1 antibody (ab56777) at 1ug/lane + NIH/3T3 cell lysate at 25 μg/lane.

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-ALDH1L1 antibody [3E9] - Astrocyte Marker (ab56777)

Predicted band size: 99 kDa

false