Anti-IL-12RB1 antibody [EPR28354-118] - BSA and Azide free

• IP

Lab

Immunoprecipitation - Anti-IL-12RB1 antibody [EPR28354-118] - BSA and Azide free (AB325507)

This data was developed using ab325502, the same antibody clone in a different buffer formulation.

IL-12RB1 was immunoprecipitated from 0.35 mg HDLM-2 (human Hodgkin lymphoma cell) whole cell lysate with ab325502 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab325502 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HDLM-2 (human Hodgkin lymphoma cell) whole cell lysate
Lane 2 : ab325502 IP in HDLM-2 (human Hodgkin lymphoma cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab325502 in HDLM-2 whole cell lysate

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

Exposure time : 180 seconds.

All lanes:

Immunoprecipitation - Anti-IL-12RB1 antibody [EPR28354-118] (<a href='/ko/products/primary-antibodies/il-12rb1-antibody-epr28354-118-ab325502'>ab325502</a>)

Lane 1:

HDLM-2 (human Hodgkin lymphoma cell) whole cell lysate at 10 µg

Lane 2:

<a href='/ko/products/primary-antibodies/il-12rb1-antibody-epr28354-118-ab325502'>ab325502</a> at 1/30 IP in HDLM-2 (human Hodgkin lymphoma cell) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/ko/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/ko/products/primary-antibodies/il-12rb1-antibody-epr28354-118-ab325502'>ab325502</a> in HDLM-2 whole cell lysate

Secondary

All lanes:

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

Observed band size: 100 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-IL-12RB1 antibody [EPR28354-118] - BSA and Azide free (AB325507)

This data was developed using ab325502, 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 aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-IL-12RB1 antibody [EPR28354-118] (<a href='/ko/products/primary-antibodies/il-12rb1-antibody-epr28354-118-ab325502'>ab325502</a>)

All lanes:

Human tonsil tissue lysate at 50 µg

Secondary

All lanes:

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

Observed band size: 100 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-IL-12RB1 antibody [EPR28354-118] - BSA and Azide free (AB325507)

This data was developed using ab325502, the same antibody clone in a different buffer formulation.

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

Low expression : SH-SY5Y, MCF7.

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-IL-12RB1 antibody [EPR28354-118] (<a href='/ko/products/primary-antibodies/il-12rb1-antibody-epr28354-118-ab325502'>ab325502</a>)

Lane 1:

KARPAS-299 (human T cell lymphoma cell) whole cell lysate at 50 µg

Lane 2:

Ramos (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 50 µg

Lane 3:

HDLM-2 (human Hodgkin lymphoma cell) whole cell lysate at 50 µg

Lane 4:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 50 µg

Lane 5:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 100 kDa,36 kDa

true

Exposure time: 81s

• WB

Lab

Western blot - Anti-IL-12RB1 antibody [EPR28354-118] - BSA and Azide free (AB325507)

This data was developed using ab325502, 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 aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-IL-12RB1 antibody [EPR28354-118] (<a href='/ko/products/primary-antibodies/il-12rb1-antibody-epr28354-118-ab325502'>ab325502</a>)

Lane 1:

THP-1 (human monocytic leukemia monocyte) transfected with scrambled siRNA control whole cell lysate at 50 µg

Lane 2:

THP-1 transfected with siRNA specifically targeting IL-12RB1 whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 100 kDa,36 kDa

true

Exposure time: 81s

• WB

Lab

Western blot - Anti-IL-12RB1 antibody [EPR28354-118] - BSA and Azide free (AB325507)

This data was developed using ab325502, 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 aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-IL-12RB1 antibody [EPR28354-118] (<a href='/ko/products/primary-antibodies/il-12rb1-antibody-epr28354-118-ab325502'>ab325502</a>)

Lane 1:

HDLM-2 (human Hodgkin lymphoma cell) transfected with scrambled siRNA control whole cell lysate at 50 µg

Lane 2:

HDLM-2 transfected with siRNA specifically targeting IL-12RB1 whole cell lysate at 50 µg

Secondary

All lanes:

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

Observed band size: 100 kDa,36 kDa

true

Exposure time: 125s