Anti-SMC1A antibody [EPR30532-7]

• ChIP-seq

Lab

ChIP-sequencing - Anti-SMC1A antibody [EPR30532-7] (AB325922)

Chromatin was prepared from MCF7 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab325922 [EPR30532-7]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-SMC1A antibody [EPR30532-7] (AB325922)

Chromatin was prepared from MCF7 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab325922 [EPR30532-7]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-SMC1A antibody [EPR30532-7] (AB325922)

Chromatin was prepared from MCF7 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab325922 [EPR30532-7]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• IP

Lab

Immunoprecipitation - Anti-SMC1A antibody [EPR30532-7] (AB325922)

SMC1A was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate with ab325922 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab325922 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab325922 IP in MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab325922 in MCF7 whole cell lysate

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

Exposure time : 15 seconds.

All lanes:

Immunoprecipitation - Anti-SMC1A antibody [EPR30532-7] (ab325922) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Lane 2:

ab325922 at 1/30 IP in MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab325922 in MCF7 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

Observed band size: 160 kDa

false

Exposure time: 15s

• IP

Lab

Immunoprecipitation - Anti-SMC1A antibody [EPR30532-7] (AB325922)

SMC1A was immunoprecipitated from 0.35 mg 3T3-L1(mouse embryonic fibroblast) whole cell lysate with ab325922 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab325922 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : 3T3-L1(mouse embryonic fibroblast) whole cell lysate
Lane 2 : ab325922 IP in 3T3-L1(mouse embryonic fibroblast) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab325922 in 3T3-L1 whole cell lysate

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

Exposure time : 180 seconds.

All lanes:

Immunoprecipitation - Anti-SMC1A antibody [EPR30532-7] (ab325922) at 1/1000 dilution

Lane 1:

3T3-L1(mouse embryonic fibroblast) whole cell lysate at 10 µg

Lane 2:

ab325922 at 1/30 IP in 3T3-L1(mouse embryonic fibroblast) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab325922 in 3T3-L1 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

Observed band size: 160 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-SMC1A antibody [EPR30532-7] (AB325922)

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

Lanes 1-2 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 and lanes 3-4 are incubated with Goat Anti-Rabbit IgG (HRP) at 1/2000.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

Exposure time : Lanes 1 and 3-4 : 180 seconds; Lane 2 : 26 seconds

All lanes:

Western blot - Anti-SMC1A antibody [EPR30532-7] (ab325922) at 1/1000 dilution

Lane 1:

Human spleen tissue lysate at 20 µg

Lane 2:

Human tonsil tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Mouse liver tissue lysate at 20 µg

Secondary

Lanes 1 - 2:

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

Lanes 3 - 4:

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

Observed band size: 160 kDa,124 kDa

false

• WB

Lab

Western blot - Anti-SMC1A antibody [EPR30532-7] (AB325922)

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

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

Exposure time : Lanes 1-2 : 26 seconds; Lanes 3-8 : 59 seconds

All lanes:

Western blot - Anti-SMC1A antibody [EPR30532-7] (ab325922) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with scrambled siRNA control fresh whole cell lysate at 20 µg

Lane 2:

293T transfected with siRNA specifically targeting SMC1A fresh whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 5:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 7:

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

Lane 8:

3T3-L1 (mouse embryonic fibroblast) 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: 160 kDa,124 kDa

false

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-SMC1A antibody [EPR30532-7] (AB325922)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (human cervical adenocarcinoma epithelial cell) cells and 5 µg of ab325922 [EPR30532-7]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-SMC1A antibody [EPR30532-7] (AB325922)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (human cervical adenocarcinoma epithelial cell) cells and 5 µg of ab325922 [EPR30532-7]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-SMC1A antibody [EPR30532-7] (AB325922)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (human cervical adenocarcinoma epithelial cell) cells and 5 µg of ab325922 [EPR30532-7]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• I-ELISA

Lab

Indirect ELISA - Anti-SMC1A antibody [EPR30532-7] (AB325922)

Indirect ELISA analysis of ab325922 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution dilution.

Antigen : Human SMC1A protein;Human SMC1B protein.

Antigen concentration : 1000 ng/ml

This antibody does not cross-react with Human SMC1B protein by ELISA.