Anti-3-Nitrotyrosine antibody [EPR27217-88]

• IP

Supplier Data

Immunoprecipitation - Anti-3-Nitrotyrosine antibody [EPR27217-88] (AB314438)

3-Nitrotyrosine was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) treated with 3 mM peroxynitrite for 30 minutes whole cell lysate with ab314438 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314438 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) treated with 3 mM peroxynitrite for 30 minutes whole cell lysate Lane 2 : ab314438 IP in HeLa (human cervix adenocarcinoma epithelial cell) treated with 3 mM peroxynitrite for 30 minutes whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314438 in HeLa treated with 3 mM peroxynitrite for 30 minutes whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-3-Nitrotyrosine antibody [EPR27217-88] (ab314438) at 1/30 dilution

All lanes:

HeLa (human cervix adenocarcinoma epithelial cell) treated with 3 mM peroxynitrite for 30 minutes 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: 15s

• WB

Supplier Data

Western blot - Anti-3-Nitrotyrosine antibody [EPR27217-88] (AB314438)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The identity of the bands around 50 kDa are unknown. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution. In Western blot, Anti-phosphotyrosine antibody [EPR16871] (ab179530) staining at 1/1000 dilution

All lanes:

Western blot - Anti-3-Nitrotyrosine antibody [EPR27217-88] (ab314438) at 1/1000 dilution

Lanes 1 and 7:

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

Lane 2:

HeLa treated with 3 mM peroxynitrite for 30 minutes whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

NIH/3T3 treated with 3 mM peroxynitrite for 30 minutes whole cell lysate at 20 µg

Lane 5:

Untreated PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

Lane 6:

PC-12 treated with 3 mM peroxynitrite for 30 minutes whole cell lysate at 20 µg

Lane 8:

HeLa treated with 1 mM pervanadate for 30 minutes 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: 1-250 kDa

false

Exposure time: 37s

• I-ELISA

Supplier Data

Indirect ELISA - Anti-3-Nitrotyrosine antibody [EPR27217-88] (AB314438)

Indirect ELISA analysis of ab314438 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution.
Antigen : 3-Nitrotyrosine peptide，BSA-crosslinked single 3-Nitrotyrosine amino acid.
Antigen concentration : 1000 ng/ml

• Dot

Supplier Data

Dot Blot - Anti-3-Nitrotyrosine antibody [EPR27217-88] (AB314438)

Dot blot analysis of 3-Nitrotyrosine using ab314438 at 1 : 1000 (0.497 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution. Exposure time : 10 seconds Blocking and diluting buffer and concentration : 5% NFDM/TBST