Anti-FSTL1/FRP antibody [EPR21933-155]

• IP

Unknown

Immunoprecipitation - Anti-FSTL1/FRP antibody [EPR21933-155] (AB223287)

FSTL1/FRP was immunoprecipitated from 0.35 mg rat embryo lysate with ab223287 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab223287 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1 : Rat embryo lysate 10 μg (Input).
Lane 2 : ab223287 IP in rat embryo lysate (+).
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab223287 in rat embryo lysate (-).

Blocking/Dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 1 minute.

The molecular mass observed is consistent with what has been described in the literature (PMID : 28574994).

All lanes:

Immunoprecipitation - Anti-FSTL1/FRP antibody [EPR21933-155] (ab223287)

Predicted band size: 35 kDa

Observed band size: 43-55 kDa

false

• WB

Unknown

Western blot - Anti-FSTL1/FRP antibody [EPR21933-155] (AB223287)

Blocking/diluting buffer and concentration : 5% NFDM/TBST.

Exposure times :
Lane 1 and 2 : 3 minutes;
Lane 3 : 15 seconds.

The molecular mass observed is consistent with what has been described in the literature (PMID : 27234440).

All lanes:

Western blot - Anti-FSTL1/FRP antibody [EPR21933-155] (ab223287) at 1/1000 dilution

Lane 1:

Human serum at 20 µg

Lane 2:

Mouse serum at 20 µg

Lane 3:

Rat serum at 20 µg

Secondary

Lanes 1 - 2:

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

Lane 3:

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: 35 kDa

Observed band size: 40-50 kDa

false

• WB

Lab

Western blot - Anti-FSTL1/FRP antibody [EPR21933-155] (AB223287)

Western blot : Anti-FSTL1/FRP antibody [EPR21933-155] ab223287 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 40 kDa in Wild-type HeLa cell lysates with no signal observed at this size in FSTL1 knockout HeLa cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc 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. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-FSTL1/FRP antibody [EPR21933-155] (ab223287) at 1/1000 dilution

Lane 1:

Wild-type HeLa at 20 µg

Lane 2:

Human FSTL1 knockout HeLa cell line at 20 µg

Lane 3:

A431 at 20 µg

Lane 4:

Ramos at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 40 kDa

Observed band size: 40 kDa

false

• WB

Unknown

Western blot - Anti-FSTL1/FRP antibody [EPR21933-155] (AB223287)

Blocking/diluting buffer and concentration : 5% NFDM/TBST.

The molecular mass observed is consistent with what has been described in the literature (PMID : 28574994).

Negative control : Rat liver (PMID : 20861081).

All lanes:

Western blot - Anti-FSTL1/FRP antibody [EPR21933-155] (ab223287) at 1/1000 dilution

Lane 1:

Rat E14 embryo lysate at 20 µg

Lane 2:

Mouse E14 embryo lysate at 20 µg

Lane 3:

C6 (Rat glial tumor cell line) cell lysate at 20 µg

Lane 4:

Rat liver 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: 35 kDa

Observed band size: 43-45 kDa

false

Exposure time: 48s