Immunohistochemical analysis of 4% paraformaldehyde-fixed mouse small cell lung cancer section labeling MASH1/Achaete-scute homolog 1 with ab211327 at 1/100 dilution, followed by Donkey anti Rabbit Alexa-594 at 1/500 dilution (red). 

Nuclear counterstain: DAPI (blue).

Positive staining of MASH1/Achaete-scute homolog 1 on mouse small cell lung cancer.

The image was kindly provided by our collaborator Dr. Hai Song, Zhejiang University.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Immunohistochemical analysis of 4% paraformaldehyde-fixed Ascl1^CreER/+; ROSA26^mTmG/+ mouse thyroid tissue labeling MASH1/Achaete-scute homolog 1 with ab211327 at 1/100 dilution, followed by Tyramide Signal Amplification (red). 

Counterstain: DAPI (blue) and chick anti-GFP (green).

ROSA26^mTmG/+ mice were bred with Ascl1^CreER/+ mice, in which the Ascl1 promoter drives the expression of Cre, to generate mice carrying the genotype of Ascl1^CreER/+; ROSA26^mTmG/+. Cre activation upon Tamoxifen administration resulted in eGFP expression from the ROSA26^mTmG/+ allele. The thyroid tissue from Ascl1^CreER/+; ROSA26^mTmG/+ mouse was fixed and embedded in paraffin. Paraffin section was stained with rabbit anti-MASH1/Achaete-scute homolog 1 (ab211327) antibody (Red color) and chick anti-GFP antibody (Green color). Tyramide signal amplification was used for high-resolution imaging of low-abundance targets.

The image was kindly provided by our collaborator Dr. Hai Song, Zhejiang University.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

All lanes : Anti-MASH1/Achaete-scute homolog 1 antibody [EPR19840] (ab211327) at 1/1000 dilution

Lane 1 : Mouse medullary thyroid cancer lysate at 20 µg
Lane 2 : NCI-H69 (Human small cell lung cancer cell line) whole cell lysate at 10 µg

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 25 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1: 15 seconds; Lane 2: 10 seconds

Ascl1 is highly expressed in medullary thyroid cancer and small cell lung cancer (PMID: 17090547 and 25267614). Our results showed specific band on mouse medullary thyroid cancer lysate and NCI-H69 (Human small cell lung cancer) lysate. The lysates were kindly provided by our collaborator Dr. Hai Song, Zhejiang University.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.5% Triton X-100 permeabilized NCI-H69 Human small cell lung cancer cells labeling MASH1/Achaete-scute homolog 1 with ab211327 at 1/100 dilution (red).

The nuclear counterstain is DAPI (blue).

Nuclear MASH1/Achaete-scute homolog 1 staining on NCI-H69 cells.

The images were kindly provided by our collaborator Dr. Hai Song, Zhejiang University.

MASH1/Achaete-scute homolog 1 was immunoprecipitated from 0.35 mg of mouse medullary thyroid cancer lysate  with ab211327 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab211327 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

Lane 1: Mouse medullary thyroid cancer lysate 10 μg (Input).

Lane 2: ab211327 IP in mouse medullary thyroid cancer lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab211327 in mouse medullary thyroid cancer lysate.

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

Exposure time: 3 minutes.

MASH1/Achaete-scute homolog 1 was immunoprecipitated from 0.35 mg of NCI-H69 (Human small cell lung cancer cell line) whole cell lysate  with ab211327 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab211327 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

Lane 1: NCI-H69 whole cell lysate 10 μg (Input).

Lane 2: ab211327 IP in NCI-H69 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab211327 in NCI-H69 whole cell lysate.

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

Exposure time: 3 minutes.