Anti-PGC1 alpha + beta antibody

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PGC1 alpha + beta antibody (AB72230)

Immunofluorescence analysis of 4% paraformaldehyde fixed HeLa cells (Human epithelial cells from cervix adenocarcinoma) labeling PGC1 alpha+beta with ab72230 at a 1/100 dilution showing nuclear staining. Permeabilisation using 0.1% tritonX-100.

Secondary ab : Anti-Rabbit Alexa Fluor® 488 (ab150077) at 1/500 dilution. Counter stain is labeling tubulin (red) with ab7291 at 1/1000 dilution with secondary antibody Anti-Mouse AlexaFluor® 594 (ab150120) at 1/500 dilution. DAPI stains the nucleus in blue. -ve control 1 is ab72230 at 1/100 dilution, Anti-Mouse AlexaFluor® 594 (ab150120) at 1/500 dilution. -ve control 2 is ab7291 at 1/1000 dilution, Anti-Rabbit Alexa Fluor® 488 (ab150077) at 1/500 dilution.

• WB

Project6386****

Western blot - Anti-PGC1 alpha + beta antibody (AB72230)

We have occasionally observed bands in the region of 90 kDa in batches of this antibody, which corresponds to the molecular weight based on primary sequence.

All lanes:

Western blot - Anti-PGC1 alpha + beta antibody (ab72230) at 1 µg/mL

Lane 1:

Testis (Mouse) Tissue Lysate at 10 µg

Lane 2:

Human skeletal muscle tissue lysate - total protein (<a href='/en-us/products/unavailable/human-skeletal-muscle-tissue-lysate-total-protein-ab29330'>ab29330</a>) at 10 µg

Lane 3:

Ovary (Rat) Tissue Lysate at 10 µg

Lane 4:

Colon (Mouse) Tissue Lysate at 10 µg

Lane 5:

Skeletal Muscle (Mouse) Tissue Lysate at 10 µg

Lane 6:

Thymus (Rat) Tissue Lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 91 kDa

Observed band size: 110 kDa,35 kDa,45 kDa,55 kDa

true

Exposure time: 2min

• ELISA

Lab

ELISA - Anti-PGC1 alpha + beta antibody (AB72230)

ab72230 was tested using an Indirect ELISA approach. The wells were coated with peptide (1μg/ml at 100μl/well) overnight at 4°C, followed by a 5% BSA blocking step for 1 hour at room temperature. The primary Ab was then added at a dilution range of 1- 0.00025μg/ml (100μl/well) for 1hr at room temperature. A HRP-conjugated anti-rabbit IgG (heavy and light chain) was used as a secondary antibody at 1 : 20,000 dilution for 1hr at room temperature.

ab72230 gave a positive result in ELISA against PGC1 alpha peptide and PGC1 beta peptide. This indicates that it recognises both forms.

Orange line : ELISA against immunising peptide ab73600
Yellow line : ELISA against ab88232

• WB

Unknown

Western blot - Anti-PGC1 alpha + beta antibody (AB72230)

Western Blot showing equal blocking with the PGC1 alpha and PGC1 beta peptides.

All lanes:

Western blot - Anti-PGC1 alpha + beta antibody (ab72230) at 1 µg/mL

Lane 1:

Human skeletal muscle tissue lysate - total protein (<a href='/en-us/products/unavailable/human-skeletal-muscle-tissue-lysate-total-protein-ab29330'>ab29330</a>) at 10 µg

Lane 2:

Colon (Mouse) Tissue Lysate at 10 µg

Lane 3:

Thymus (Rat) Tissue Lysate at 10 µg

Lane 4:

Human skeletal muscle tissue lysate - total protein (<a href='/en-us/products/unavailable/human-skeletal-muscle-tissue-lysate-total-protein-ab29330'>ab29330</a>) at 10 µg with Human PGC1 alpha peptide (<a href='/en-us/products/unavailable/human-pgc1-alpha-peptide-ab73600'>ab73600</a>)

Lane 5:

Colon (Mouse) Tissue Lysate at 10 µg with Human PGC1 alpha peptide (<a href='/en-us/products/unavailable/human-pgc1-alpha-peptide-ab73600'>ab73600</a>)

Lane 6:

Thymus (Rat) Tissue Lysate at 10 µg with Human PGC1 alpha peptide (<a href='/en-us/products/unavailable/human-pgc1-alpha-peptide-ab73600'>ab73600</a>)

Lane 7:

Human skeletal muscle tissue lysate - total protein (<a href='/en-us/products/unavailable/human-skeletal-muscle-tissue-lysate-total-protein-ab29330'>ab29330</a>) at 10 µg with PGC1 beta peptide

Lane 8:

Colon (Mouse) Tissue Lysate at 10 µg with PGC1 beta peptide

Lane 9:

Thymus (Rat) Tissue Lysate at 10 µg with PGC1 beta peptide

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 91 kDa

Observed band size: 110 kDa,45 kDa,73 kDa,90 kDa

false

Exposure time: 8min