Anti-Osteocalcin antibody [EPR24665-63] - BSA and Azide free

• WB

Lab

Western blot - Anti-Osteocalcin antibody [EPR24665-63] - BSA and Azide free (AB309522)

Western blot : Anti-OCN antibody [EPR24665-63] (ab309521) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab309521 was shown to bind specifically to OCN. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged after ECL incubation followed by 20 minute exposure. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Osteocalcin antibody [EPR24665-63] (<a href='/en-us/products/primary-antibodies/osteocalcin-antibody-epr24665-63-ab309521'>ab309521</a>) at 1/1000 dilution

Lane 1:

MC3T3-E1 undifferentiated whole lysate cell lysate at 20 µg

Lane 2:

MC3T3-E1 7-day Osteogenic differentiation Whole Lysate cell lysate at 20 µg

Lane 3:

MC3T3-E1 14-day Osteogenic differentiation Whole Lysate cell lysate at 20 µg

Lane 4:

MC3T3-E1 28-day Osteogenic differentiation Whole Lysate cell lysate at 20 µg

Lane 5:

NIH/3T3 cell lysate at 20 µg

Lane 6:

C2C12 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>)

false

• WB

Supplier Data

Western blot - Anti-Osteocalcin antibody [EPR24665-63] - BSA and Azide free (AB309522)

This data was developed using ab309521, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : mouse brain (PMID : 7575621), mouse lung (PMID : 7575621). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-Osteocalcin antibody [EPR24665-63] (<a href='/en-us/products/primary-antibodies/osteocalcin-antibody-epr24665-63-ab309521'>ab309521</a>) at 1/1000 dilution

Lane 1:

Mouse articar cartilage tissue lysate 50 at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Mouse lu tissue lysate 50 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 1/20000 dilution

Observed band size: 15 kDa

false

Exposure time: 180s

• Dot

Supplier Data

Dot Blot - Anti-Osteocalcin antibody [EPR24665-63] - BSA and Azide free (AB309522)

This data was developed using ab309521, the same antibody clone in a different buffer formulation. Dot blot analysis of Osteocalcin using ab309521 at 1 : 1000 (0.516 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution. Lane 1 : Mouse Bglap1 recombinant protein Lane 2 : Mouse Bglap3 recombinant protein Lane 3 : Mouse Bglap2 recombinant protein Exposure time : 37 seconds Blocking and diluting buffer and concentration : 5% NFDM/TBST

• WB

Supplier Data

Western blot - Anti-Osteocalcin antibody [EPR24665-63] - BSA and Azide free (AB309522)

This data was developed using ab309521, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST In Western blot, anti- H3 antibody (ab176842) loading control staining at 1/100000 dilution. Anti-His antibody (ab213204) staining at 1/5000 dilution. Exposure time : 48 seconds

All lanes:

Western blot - Anti-Osteocalcin antibody [EPR24665-63] (<a href='/en-us/products/primary-antibodies/osteocalcin-antibody-epr24665-63-ab309521'>ab309521</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containi a his tag whole cell lysate 20 at 20 µg

Lane 2:

293T cells transfected with a mosue Osteocalcin expression vector containi a His-tag whole cell lysate 20 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

Observed band size: 15 kDa

false

Exposure time: 48s