Anti-LZTR1 antibody [EPR25253-51] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-LZTR1 antibody [EPR25253-51] - BSA and Azide free (AB289988)

This data was developed using ab289965, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling LZTR1 with ab289965 at 1/50 dilution (1μg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-LZTR1 antibody [EPR25253-51] - BSA and Azide free (AB289988)

This data was developed using ab289965, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling LZTR1 with ab289965 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluo^r® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing cytoplasm staining in SH-SY5Y cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluo^r® 488) preadsorbed at 1/1000 dilution.

• IP

Lab

Immunoprecipitation - Anti-LZTR1 antibody [EPR25253-51] - BSA and Azide free (AB289988)

This data was developed using ab289965, the same antibody clone in a different buffer formulation.

LZTR1 was immunoprecipitated from HEK-293T ( human embryonic kidney epithelial cell) whole cell lysate with ab289965 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab289965 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HEK-293T ( human embryonic kidney epithelial cell) whole cell lysate 10 μg

Lane 2 : ab289965 IP in HEK-293T whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab289965 in HEK-293T whole cell lysate

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

Exposure time : 41 seconds

All lanes:

Immunoprecipitation - Anti-LZTR1 antibody [EPR25253-51] (<a href='/en-us/products/primary-antibodies/lztr1-antibody-epr25253-51-ab289965'>ab289965</a>)

Predicted band size: 94 kDa

Observed band size: 95 kDa

false

• IP

Lab

Immunoprecipitation - Anti-LZTR1 antibody [EPR25253-51] - BSA and Azide free (AB289988)

This data was developed using ab289965, the same antibody clone in a different buffer formulation.

LZTR1 was immunoprecipitated from NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab289965 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab289965 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 lysate10 μg

Lane 2 : ab289965 IP in NIH/3T3 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab289965 in NIH/3T3 whole cell lysate

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

Exposure time : 41 seconds

All lanes:

Immunoprecipitation - Anti-LZTR1 antibody [EPR25253-51] (<a href='/en-us/products/primary-antibodies/lztr1-antibody-epr25253-51-ab289965'>ab289965</a>)

Predicted band size: 94 kDa

Observed band size: 95 kDa

false

• WB

Lab

Western blot - Anti-LZTR1 antibody [EPR25253-51] - BSA and Azide free (AB289988)

This data was developed using ab289965, the same antibody clone in a different buffer formulation.

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

The bands nearby 50 kDa could be non-specific bands.

Exposure times : Lane 1 : 70 seconds; Lanes 2-3 : 180 seconds.

All lanes:

Western blot - Anti-LZTR1 antibody [EPR25253-51] (<a href='/en-us/products/primary-antibodies/lztr1-antibody-epr25253-51-ab289965'>ab289965</a>) at 1/1000 dilution

Lane 1:

K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 2:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell 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 100000 Cells

Lanes 1 - 3:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/100000 dilution

Predicted band size: 94 kDa

Observed band size: 95 kDa

false

• WB

Lab

Western blot - Anti-LZTR1 antibody [EPR25253-51] - BSA and Azide free (AB289988)

This data was developed using ab289965, the same antibody clone in a different buffer formulation.

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

The bands nearby 50 kDa could be non-specific bands.

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-LZTR1 antibody [EPR25253-51] (<a href='/en-us/products/primary-antibodies/lztr1-antibody-epr25253-51-ab289965'>ab289965</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 12 µg

Lane 2:

HeLa transfected with siRNA specifically targeting LZTR1 whole cell lysate at 12 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 1/100000 dilution

Predicted band size: 94 kDa

Observed band size: 95 kDa

false