Anti-NLRX1 antibody [EPR27216-40]

• IP

Supplier Data

Immunoprecipitation - Anti-NLRX1 antibody [EPR27216-40] (AB307668)

NLRX1 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate 10 ug with ab307668 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307668 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate 10 ug
Lane 2 : ab307668 IP in HeLa whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307668 in HeLa whole cell lysate

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

Exposure time : 180 seconds

All lanes:

Immunoprecipitation - Anti-NLRX1 antibody [EPR27216-40] (ab307668) at 1/1000 dilution

All lanes:

HeLa cell

Secondary

All lanes:

Immunoprecipitation at 1/5000 dilution

Observed band size: 100 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-NLRX1 antibody [EPR27216-40] (AB307668)

Blocking and diluting buffer and concentration : ½ volume of Odyssey Blocking Buffer (TBS)+ ½ volume of 0.1% TBS The samples were run on a Bis-Tris gel. Performed under reducing conditions. False colour image of Western blot : Anti-NLRX1 antibody [EPR27216-40] (ab307668) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab307668 was shown to bind specifically to NLRX1. A band was observed at 100 kDa in wild-type HeLa cell lysates whereas no signal observed at this size in NLRX1 knockout cell line ab265964 (knockout cell lysate ab258074). To generate this image, wild-type and NLRX1 knockout HeLa cell lysates were analyzed. First, samples were run on a SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS 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 IgG H&L (IRDye® 800CW) preabsorbed (ab216773) at 1/10000 dilution and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

Lane 1:

Wild-type HeLa whole cell lysate at 20 µg

Lane 2:

NLRX1 knockout HeLa whole cell lysate at 20 µg

Secondary

All lanes:

Western blot at 1/10000 dilution

Observed band size: 100 kDa

false

• WB

Supplier Data

Western blot - Anti-NLRX1 antibody [EPR27216-40] (AB307668)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Primary antibody staining at 1/1000 dilution and the secondary antibody, Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051), staining at 1/20000 dilution Exposure time : 59 seconds

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Anti-NLRX1 antibody [EPR27216-40] (ab307668) at 1/20000 dilution

Observed band size: 100 kDa

false

Exposure time: 59s