Anti-MASH1/Achaete-scute homolog 1 antibody [RM1182]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MASH1/Achaete-scute homolog 1 antibody [RM1182] (AB322665)

Immunohistochemical analysis of paraffin-embedded Human thyroid carcinoma tissue labeling MASH1/Achaete-scute homolog 1 with ab322665 at 1/50 (10.42 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on human thyroid carcinoma. The section was incubated with ab322665 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MASH1/Achaete-scute homolog 1 antibody [RM1182] (AB322665)

Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling MASH1/Achaete-scute homolog 1 with ab322665 at 1/50 (10.42 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on human lung carcinoma. The section was incubated with ab322665 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MASH1/Achaete-scute homolog 1 antibody [RM1182] (AB322665)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling MASH1/Achaete-scute homolog 1 with ab322665 at 1/50 (10.42 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Negative control : no staining on human kidney. The section was incubated with ab322665 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-MASH1/Achaete-scute homolog 1 antibody [RM1182] (AB322665)

Chromatin was prepared from NCI-H69 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 µg of ab322665 [RM1182]. 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-MASH1/Achaete-scute homolog 1 antibody [RM1182] (AB322665)

Chromatin was prepared from NCI-H69 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 µg of ab322665 [RM1182]. 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-MASH1/Achaete-scute homolog 1 antibody [RM1182] (AB322665)

MASH1/Achaete-scute homolog 1 was immunoprecipitated from 0.35 mg NCI-H69 (human lung small cell carcinoma cell line) whole cell lysate with ab322665 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322665 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : NCI-H69 (human lung small cell carcinoma cell line) whole cell lysate
Lane 2 : ab322665 IP in NCI-H69 (human lung small cell carcinoma cell line) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab322665 in NCI-H69 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-MASH1/Achaete-scute homolog 1 antibody [RM1182] (ab322665) at 1/30 dilution

All lanes:

NCI-H69 (human lung small cell carcinoma cell line) 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: 76s

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-MASH1/Achaete-scute homolog 1 antibody [RM1182] (AB322665)

Chromatin was prepared from NCI-H69 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 µg of ab322665 [RM1182]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MASH1/Achaete-scute homolog 1 antibody [RM1182] (AB322665)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling MASH1/Achaete-scute homolog 1 with ab322665 at 1/100 (5.21 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on rat cerebrum. The section was incubated with ab322665 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-MASH1/Achaete-scute homolog 1 antibody [RM1182] (AB322665)

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

In lane 2, the lysate was stored at -80℃ prior to Western Blotting. The bands beneath the target band (30 kDa) are likey to be degradation products. In lane 1/3, To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

Exposure time : Lanes 1-2 : 180 seconds; Lane 3 : 59 seconds

All lanes:

Western blot - Anti-MASH1/Achaete-scute homolog 1 antibody [RM1182] (ab322665) at 1/1000 dilution

Lanes 1 - 2:

NCI-H69 (human lung small cell carcinoma cell line) whole cell lysate at 20 µg

Lane 3:

SH-SY5Y (human neuroblastoma 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 1/20000 dilution

Observed band size: 30 kDa

false

• WB

Supplier Data

Western blot - Anti-MASH1/Achaete-scute homolog 1 antibody [RM1182] (AB322665)

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

Negative control : A549 (PMID : 31231131).

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

All lanes:

Western blot - Anti-MASH1/Achaete-scute homolog 1 antibody [RM1182] (ab322665) at 1/1000 dilution

Lane 1:

NCI-H69 (human lung small cell carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

A549 (human lung carcinoma 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 1/20000 dilution

Observed band size: 30 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-MASH1/Achaete-scute homolog 1 antibody [RM1182] (AB322665)

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

Negative control : kidney

The bands beneath the target band (30 kDa) are likey to be degradation products.

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

All lanes:

Western blot - Anti-MASH1/Achaete-scute homolog 1 antibody [RM1182] (ab322665) at 1/1000 dilution

Lane 1:

Human brain tissue lysate at 20 µg

Lane 2:

Human kidney tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse kidney tissue lysate at 20 µg

Lane 5:

Rat brain tissue lysate at 20 µg

Lane 6:

Rat kidney 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: 30 kDa,36 kDa

false

Exposure time: 180s