Anti-HOXA10 antibody [EPR27317-76]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-HOXA10 antibody [EPR27317-76] (AB308516)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized K-562 (human chronic myelogenous leukemia lymphoblast, Left) / THP-1 (human monocytic leukemia monocyte, Right) cells labelling HOXA10 with ab308516 at 1/50 dilution (1 ug)/Red (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 1/5000 dilution was used as the secondary antibody. Negative control : K-562 (PMID : 10766757).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-HOXA10 antibody [EPR27317-76] (AB308516)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized THP-1 (human monocytic leukemia monocyte) cells labelling HOXA10 with ab308516 at 1/50 (9.52 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing nuclear and weak cytoplasmic staining in THP-1 cell line.Negative control : K-562 (PMID 10766757).Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ChIP-seq

Lab

ChIP-sequencing - Anti-HOXA10 antibody [EPR27317-76] (AB308516)

Chromatin was prepared from THP-1 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab308516 [EPR27317-76]. 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-HOXA10 antibody [EPR27317-76] (AB308516)

Chromatin was prepared from THP-1 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab308516 [EPR27317-76]. 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-HOXA10 antibody [EPR27317-76] (AB308516)

Chromatin was prepared from THP-1 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab308516 [EPR27317-76]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• IP

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Immunoprecipitation - Anti-HOXA10 antibody [EPR27317-76] (AB308516)

HOXA10 was immunoprecipitated from 0.35 mg THP-1 whole cell lysate with ab308516 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab308516 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : THP-1 (human monocytic leukemia monocyte) whole cell lysate Lane 2 : ab308516 IP in THP-1 whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab308516 in THP-1 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 32 seconds

All lanes:

Immunoprecipitation - Anti-HOXA10 antibody [EPR27317-76] (ab308516) at 1/30 dilution

All lanes:

THP-1 (human monocytic leukemia monocyte) 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: 32s

• WB

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Western blot - Anti-HOXA10 antibody [EPR27317-76] (AB308516)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : K-562(PMID : 7527557) Bands below 50 kDa may due to degradation. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 114 seconds

All lanes:

Western blot - Anti-HOXA10 antibody [EPR27317-76] (ab308516) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

K562 (human chronic myelogenous leukemia lymphoblast) 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: 50 kDa

false

Exposure time: 114s

• WB

Supplier Data

Western blot - Anti-HOXA10 antibody [EPR27317-76] (AB308516)

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

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

All lanes:

Western blot - Anti-HOXA10 antibody [EPR27317-76] (ab308516) at 1/1000 dilution

Lane 1:

Human bone marrow tissue lysate at 20 µg

Lane 2:

Human testis 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 kDa

true

Exposure time: 180s

• Dot

Supplier Data

Dot Blot - Anti-HOXA10 antibody [EPR27317-76] (AB308516)

Dot blot analysis of HOXA10 using ab308516 at 1 : 1000 (0.476 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution. Lane 1 : Human Homeobox protein Hox-A10 peptide Lane 2 : Human Homeobox protein Hox-C10 peptide Lane 3 : Human Homeobox protein Hox-D10 peptide Exposure time : 180 seconds Blocking and diluting buffer and concentration : 5% NFDM/TBST

• WB

Supplier Data

Western blot - Anti-HOXA10 antibody [EPR27317-76] (AB308516)

Blocking and diluting buffer and concentration : 5% NFDM/TBST This antibody does not cross-react with human HOXA9. In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-HOXA10 antibody [EPR27317-76] (ab308516) at 1/1000 dilution

Lane 1:

His-tagged human HOXA9 recombinant protein fragment at 10 ng

Lane 2:

His-tagged human HOXA10 recombinant protein fragment at 10 ng

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: 15 kDa

false

Exposure time: 180s