Anti-KAT2A / GCN5 antibody [EPR28321-83] - BSA and Azide free

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-KAT2A / GCN5 antibody [EPR28321-83] - BSA and Azide free (AB321886)

This data was developed using ab321885, the same antibody clone in a different buffer formulation.

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 μg of ab321885 [EPR28321-83]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-KAT2A / GCN5 antibody [EPR28321-83] - BSA and Azide free (AB321886)

This data was developed using ab321885, the same antibody clone in a different buffer formulation.

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 μg of ab321885 [EPR28321-83]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-KAT2A / GCN5 antibody [EPR28321-83] - BSA and Azide free (AB321886)

This data was developed using ab321885, the same antibody clone in a different buffer formulation.

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 μg of ab321885 [EPR28321-83]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• IP

Supplier Data

Immunoprecipitation - Anti-KAT2A / GCN5 antibody [EPR28321-83] - BSA and Azide free (AB321886)

This data was developed using ab321885, the same antibody clone in a different buffer formulation.

KAT2A / GCN5 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab321885 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab321885 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
Lane 2 : ab321885 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab321885 in HeLa whole cell lysate

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

Lysates were freshly made and used for Western blotting immediately after immunoprecipitation to minimize protein degradation. The identity of the lower MW band at approximately 55 kDa are likely to be degraded target fragments.

All lanes:

Immunoprecipitation - Anti-KAT2A / GCN5 antibody [EPR28321-83] (<a href='/en-us/products/primary-antibodies/kat2a-gcn5-antibody-epr28321-83-ab321885'>ab321885</a>) at 1/30 dilution

All lanes:

HeLa (human cervical adenocarcinoma epithelial cell) 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: 58s

• WB

Supplier Data

Western blot - Anti-KAT2A / GCN5 antibody [EPR28321-83] - BSA and Azide free (AB321886)

This data was developed using ab321885, the same antibody clone in a different buffer formulation.

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

Performed under reducing conditions.

In Western blot, ab321885 was shown to bind specifically to KAT2 / GCN5. Target of interest was observed at 95 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in KAT2A / GCN5 knockout cell line (lane 2).

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

All lanes:

Western blot - Anti-KAT2A / GCN5 antibody [EPR28321-83] (<a href='/en-us/products/primary-antibodies/kat2a-gcn5-antibody-epr28321-83-ab321885'>ab321885</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 2:

KAT2A / GCN5 knockout HAP1 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 1/20000 dilution

Observed band size: 95 kDa,36 kDa

false

Exposure time: 180s

• Dot

Supplier Data

Dot Blot - Anti-KAT2A / GCN5 antibody [EPR28321-83] - BSA and Azide free (AB321886)

This data was developed using ab321885, the same antibody clone in a different buffer formulation.

Dot blot analysis of KAT2A / GCN5 using ab321885 at 1000 (0.519 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : His-tagged human KAT2A fragment
Lane2 : His-tagged human KAT2B fragment

Exposure time : 180 seconds.

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

This antibody does not cross-react with human KAT2B.

#In Dot Blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Dot Blot - Anti-KAT2A / GCN5 antibody [EPR28321-83] (<a href='/en-us/products/primary-antibodies/kat2a-gcn5-antibody-epr28321-83-ab321885'>ab321885</a>) at 1/1000 dilution

Lane 1:

His-tagged human KAT2A fragment

Lane 2:

His-tagged human KAT2B fragment

Secondary

All lanes:

Dot 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

false

Exposure time: 180s