Anti-ADAM9 antibody [EPR21924-164] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-ADAM9 antibody [EPR21924-164] - BSA and Azide free (AB238647)

ADAM9 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma cell) whole cell lysate with ab218242 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab218242 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.

Lane 1 : HeLa whole cell lysate 10μg (input).
Lane 2 : ab218242 IP in HeLa whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab218242 in HeLa whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 3 minutes.

The conformational epitope maybe changed after cleavage and this antibody has better affinity to the pro-form.

The molecular mass observed is consistent with what has been described in the literature (PMID 29118335; PMID : 12767059; PMID : 20736367).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218242).

All lanes:

Immunoprecipitation - Anti-ADAM9 antibody [EPR21924-164] (<a href='/ko/products/primary-antibodies/adam9-antibody-epr21924-164-ab218242'>ab218242</a>)

Predicted band size: 70 kDa,85 kDa,91 kDa

Observed band size: 100 kDa,75 kDa

false

• WB

Supplier Data

Western blot - Anti-ADAM9 antibody [EPR21924-164] - BSA and Azide free (AB238647)

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

Exposure times :
Lanes 1-5 : 3 minutes
Lane 6 : 48 seconds
Lane 7 : 114 seconds

The molecular mass observed is consistent with what has been described in the literature (PMID 29118335; PMID : 12767059; PMID : 20736367)

We recommend that customers avoid boiling their samples to prevent protein aggregation.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218242).

All lanes:

Western blot - Anti-ADAM9 antibody [EPR21924-164] (<a href='/ko/products/primary-antibodies/adam9-antibody-epr21924-164-ab218242'>ab218242</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 10 µg

Lane 2:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 3:

WI-38 (human fetal lung fibroblast), whole cell lysate at 10 µg

Lane 4:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 5:

U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate at 10 µg

Lane 6:

A549 (human lung carcinoma epithelial cell), whole cell lysate at 10 µg

Lane 7:

A431 (human epidermoid carcinoma epithelial cell), whole cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 91 kDa

Observed band size: 100 kDa,75 kDa

false

• WB

Lab

Western blot - Anti-ADAM9 antibody [EPR21924-164] - BSA and Azide free (AB238647)

This data was developed using the same antibody clone in a different buffer formulation (ab218242).

Lanes 1-3 : Merged signal (red and green). Green - ab218242 observed at 100,75 kDa. Red - loading control ab8245 observed at 36 kDa.

ab218242 Anti-ADAM9 antibody [EPR21924-164] was shown to specifically react with ADAM9 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265484 (knockout cell lysate ab257814) was used. Wild-type and ADAM9 knockout samples were subjected to SDS-PAGE. ab218242 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ADAM9 antibody [EPR21924-164] (<a href='/ko/products/primary-antibodies/adam9-antibody-epr21924-164-ab218242'>ab218242</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ADAM9 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ADAM9 knockout HeLa cell line (<a href='/ko/products/cell-lines/human-adam9-knockout-hela-cell-line-ab265484'>ab265484</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 91 kDa

Observed band size: 100 kDa,75 kDa

false