Anti-CD130 (gp130) antibody [EPR24557-50] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-CD130 (gp130) antibody [EPR24557-50] - BSA and Azide free (AB283713)

This data was developed using ab283685, the same antibody clone in a different buffer formulation.

CD130 (gp130) was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug with ab283685 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283685 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug

Lane 2 : ab283685 IP in NIH/3T3 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab283685 in NIH/3T3 whole cell lysate

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

Exposure time : 15 seconds

All lanes:

Immunoprecipitation - Anti-CD130 (gp130) antibody [EPR24557-50] (<a href='/en-us/products/primary-antibodies/cd130-gp130-antibody-epr24557-50-ab283685'>ab283685</a>)

Predicted band size: 104 kDa

Observed band size: 130 kDa

false

• WB

Lab

Western blot - Anti-CD130 (gp130) antibody [EPR24557-50] - BSA and Azide free (AB283713)

This data was developed using ab283685, the same antibody clone in a different buffer formulation.

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

ab283685 was shown to specifically react with IL6ST (CD130/gp130) in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266938 (knockout cell lysate ab257207) was used. Wild-type and IL6ST (CD130/gp130) knockout samples were subjected to SDS-PAGE. ab283685 and ab181602 (Rabbit anti-GAPDH loading control) were incubated for 1 hour at room temperature at 1/1000 dilution and 1/200, 000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/50, 000 dilution and 1/100, 000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.

Samples are non-boiled as boiling may cause protein aggregates.

Exposure time : 70 seconds

All lanes:

Western blot - Anti-CD130 (gp130) antibody [EPR24557-50] (<a href='/en-us/products/primary-antibodies/cd130-gp130-antibody-epr24557-50-ab283685'>ab283685</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

IL6ST (CD130/gp130) knockout A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Western blot - Human IL6ST (CD130/gp130) knockout A549 cell line (<a href='/en-us/products/cell-lines/human-il6st-cd130-gp130-knockout-a549-cell-line-ab266938'>ab266938</a>)

Lane 3:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 104 kDa

Observed band size: 130 kDa

false

• WB

Lab

Western blot - Anti-CD130 (gp130) antibody [EPR24557-50] - BSA and Azide free (AB283713)

This data was developed using ab283685, the same antibody clone in a different buffer formulation.

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

Samples are non-boiled as boiling may cause protein aggregates.

Exposure time : Lanes 1-2 : 48 seconds; Lanes 3-4 : 26 seconds.

All lanes:

Western blot - Anti-CD130 (gp130) antibody [EPR24557-50] (<a href='/en-us/products/primary-antibodies/cd130-gp130-antibody-epr24557-50-ab283685'>ab283685</a>) at 1/1000 dilution

Lane 1:

Mouse thymus tissue lysate at 20 µg

Lane 2:

Mouse lung tissue lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 whole cell lysate treated with PNGase F at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 104 kDa

Observed band size: 130 kDa

false

• WB

Lab

Western blot - Anti-CD130 (gp130) antibody [EPR24557-50] - BSA and Azide free (AB283713)

This data was developed using ab283685, the same antibody clone in a different buffer formulation.

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

gp130 is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID : 19915009).

Low expression : Daudi (PMID : 2261637).

Samples are non-boiled as boiling may cause protein aggregates.

Exposure time : Lanes 1-2 and lane 7 : 70 seconds; Lanes 3-6 : 26 seconds

All lanes:

Western blot - Anti-CD130 (gp130) antibody [EPR24557-50] (<a href='/en-us/products/primary-antibodies/cd130-gp130-antibody-epr24557-50-ab283685'>ab283685</a>) at 1/1000 dilution

Lane 1:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 3:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

MG-63 (human osteosarcoma fibroblast) whole cell lysate at 20 µg

Lane 5:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 6:

MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate at 20 µg

Lane 7:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 104 kDa

Observed band size: 130 kDa

false