Anti-IL1 Receptor I/IL-1R-1 antibody [RM1242]

• IP

Supplier Data

Immunoprecipitation - Anti-IL1 Receptor I/IL-1R-1 antibody [RM1242] (AB322149)

IL1 Receptor I/IL-1R-1 was immunoprecipitated from 0.35 mg HDLM-2 (human Hodgkin lymphoma) whole cell lysate with ab322149 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322149 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HDLM-2 (human Hodgkin lymphoma) whole cell lysate
Lane 2 : ab322149 IP in HDLM-2 (human Hodgkin lymphoma) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab322149 in HDLM-2 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-IL1 Receptor I/IL-1R-1 antibody [RM1242] (ab322149) at 1/30 dilution

All lanes:

HDLM-2 (human Hodgkin lymphoma) 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: 180s

• IP

Supplier Data

Immunoprecipitation - Anti-IL1 Receptor I/IL-1R-1 antibody [RM1242] (AB322149)

IL1 Receptor I/IL-1R-1 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab322149 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322149 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 : ab322149 IP in NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab322149 in NIH/3T3 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-IL1 Receptor I/IL-1R-1 antibody [RM1242] (ab322149) 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: 180s

• WB

Supplier Data

Western blot - Anti-IL1 Receptor I/IL-1R-1 antibody [RM1242] (AB322149)

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

Low expression : cerebellum and skeletal muscle.

The identity of the higher MW band at approximately 80 kDa is membrane-bond form of IL1R1 and the lower MW band at approximately 50 kDa is soluble(s) form of IL1R1. The bands between the 50 to 80 kDa are expected to be degradation fragments (PMID : 17324958).

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-IL1 Receptor I/IL-1R-1 antibody [RM1242] (ab322149) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human skeletal muscle tissue lysate at 20 µg

Lane 4:

Human spleen tissue 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: 50-80 kDa,36 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-IL1 Receptor I/IL-1R-1 antibody [RM1242] (AB322149)

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

Negative control : HT-1080, 293T (PMID : 18996842).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-IL1 Receptor I/IL-1R-1 antibody [RM1242] (ab322149) at 1/1000 dilution

Lane 1:

HDLM-2 (human Hodgkin lymphoma cell) whole cell lysate at 20 µg

Lane 2:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

293T (human embryonic kidney 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: 80 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-IL1 Receptor I/IL-1R-1 antibody [RM1242] (AB322149)

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

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

The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and IL1r1-KO homozygous mice (Strain ID : T005977). This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-IL1 Receptor I/IL-1R-1 antibody [RM1242] (ab322149) at 1/1000 dilution

Lane 1:

Wild-type mouse spleen tissue lysate (female) at 20 µg

Lane 2:

IL1r1 knockout mouse spleen tissue lysate (female) at 20 µg

Lane 3:

Wild-type mouse spleen tissue lysate (male) at 20 µg

Lane 4:

IL1r1 knockout mouse spleen tissue lysate (male) at 20 µg

Lane 5:

Wild-type mouse thymus tissue lysate (female) at 20 µg

Lane 6:

IL1r1 knockout mouse thymus tissue lysate (female) at 20 µg

Lane 7:

Wild-type mouse thymus tissue lysate (male) at 20 µg

Lane 8:

IL1r1 knockout mouse thymus tissue lysate (male) 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 1/20000 dilution

Observed band size: 81 kDa,36 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-IL1 Receptor I/IL-1R-1 antibody [RM1242] (AB322149)

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

Low expression : spleen, HL-1.

Negative control : B16-F10 (PMID : 29445180).

This blot of lane 5-6 was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane 1-4 : 180 seconds Lane 5-6 : 114 seconds

All lanes:

Western blot - Anti-IL1 Receptor I/IL-1R-1 antibody [RM1242] (ab322149) at 1/1000 dilution

Lane 1:

EL4.IL-2 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg

Lane 2:

HL-1 (mouse atrial muscle cell) whole cell lysate at 20 µg

Lane 3:

B16-F10 (mouse skin melanoma cell) whole cell lysate at 20 µg

Lane 4:

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

Lane 5:

Mouse spleen tissue lysate at 20 µg

Lane 6:

Mouse skeletal muscle tissue 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 1/20000 dilution

Observed band size: 50-80 kDa,36 kDa

false