Anti-NCOA4 antibody [EPR28511-12] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-NCOA4 antibody [EPR28511-12] - BSA and Azide free (AB314554)

This data was developed using ab314553, the same antibody clone in a different buffer formulation. NCOA4 was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab314553 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314553 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate Lane 2 : ab314553 IP in HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314553 in HepG2 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-NCOA4 antibody [EPR28511-12] (<a href='/en-us/products/primary-antibodies/ncoa4-antibody-epr28511-12-ab314553'>ab314553</a>) at 1/30 dilution

All lanes:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 32s

• IP

Supplier Data

Immunoprecipitation - Anti-NCOA4 antibody [EPR28511-12] - BSA and Azide free (AB314554)

This data was developed using ab314553, the same antibody clone in a different buffer formulation. NCOA4 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab314553 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314553 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 Lane 2 : ab314553 IP in NIH/3T3 (mouse embryonic fibroblast) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314553 in NIH/3T3 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-NCOA4 antibody [EPR28511-12] (<a href='/en-us/products/primary-antibodies/ncoa4-antibody-epr28511-12-ab314553'>ab314553</a>) at 1/30 dilution

All lanes:

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

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 32s

• WB

Supplier Data

Western blot - Anti-NCOA4 antibody [EPR28511-12] - BSA and Azide free (AB314554)

This data was developed using ab314553, the same antibody clone in a different buffer formulation.

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 24695223, 31061094).

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-NCOA4 antibody [EPR28511-12] (<a href='/en-us/products/primary-antibodies/ncoa4-antibody-epr28511-12-ab314553'>ab314553</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 4:

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

Lane 5:

PC-12 (rat adrenal gland pheochromocytoma) whole cell 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

Observed band size: 75 kDa,76 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-NCOA4 antibody [EPR28511-12] - BSA and Azide free (AB314554)

This data was developed using ab314553, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 24695223, 31061094). The lysates of lanes 2-3 were freshly made and used for Western blotting immediately to minimize protein degradation. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-NCOA4 antibody [EPR28511-12] (<a href='/en-us/products/primary-antibodies/ncoa4-antibody-epr28511-12-ab314553'>ab314553</a>) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Lane 3:

Rat testis tissue lysate at 20 µg

Secondary

Lanes 1 - 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

Lanes 1 - 3:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 75 kDa,76 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-NCOA4 antibody [EPR28511-12] - BSA and Azide free (AB314554)

This data was developed using ab314553, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 24695223, 31061094). Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-NCOA4 antibody [EPR28511-12] (<a href='/en-us/products/primary-antibodies/ncoa4-antibody-epr28511-12-ab314553'>ab314553</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellar carcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HepG2 transfected with siRNA specifically targeti NCOA4 whole cell 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

Observed band size: 75 kDa,76 kDa

false

Exposure time: 147s

• WB

Supplier Data

Western blot - Anti-NCOA4 antibody [EPR28511-12] - BSA and Azide free (AB314554)

This data was developed using ab314553, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : HT-1080. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 24695223, 31061094). In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-NCOA4 antibody [EPR28511-12] (<a href='/en-us/products/primary-antibodies/ncoa4-antibody-epr28511-12-ab314553'>ab314553</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

A-172 (human brain glioblastoma) whole cell lysate at 20 µg

Lane 3:

HT-1080 (human fibrosarcoma epithelial cell) whole cell 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

Observed band size: 75 kDa,76 kDa

false

Exposure time: 180s