AM32710PU-N

antibody from Acris Antibodies GmbH

Targeting: NEUROD1 BETA2, BHF-1, bHLHa3, MODY6, NEUROD

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

AM32710PU-N

Provider

Acris Antibodies GmbH

Product name

anti NEUROD1

Antibody type

Monoclonal

Antigen

Recombinant NEUROD1 protein.

Reactivity

Human, Mouse

Host

Mouse

Isotype

IgG

Antibody clone number

BC18-1.11EG

Vial size

0.1 mg

Concentration

0.5 mg/ml (Lot specific)

Western blot

Supportive validation

Submitted by

Acris Antibodies GmbH (provider)

Main image

Experimental details

Figure 3. Shows reactivity of anti-NEUROD1 Antibody Cat.-No AM32710PU-N (Clone BC18-1.11EG) against SHSY5Y, Kelly and SKNSH neuroblastoma cell lysates by Western blotting. For each cell line, a band of approximately 45kD was observed. This molecular size corresponds to the expected size for NEUROD1. The antibody was used at 2 µg/ml and incubated overnight at 4°C The peroxidase conjugated  anti-Mouse Ig secondary antibody was used at 1/10000 dilution and incubated for 2 hours. The blot was developed using enhanced chemiluminescence and image captured on radiographic films.

Immunocytochemistry

Supportive validation

Submitted by

Acris Antibodies GmbH (provider)

Main image

Experimental details

Figure 1. Shows Neuro2A (Mouse neuroblastoma) and SKNSH, 1691 and IMR32 (Human neuroblastoma) cells stained with anti-NEUROD1 Antibody Cat.-No AM32710PU-N (Clone BC18-1.11EG) by the Indirect Immunofluorescence method. The secondary antibody was FITC conjugated anti-Mouse Ig. Nuclear and cytoplasmic staining for NEUROD is observed in all cells. Since the cell lines tested loosely attach to surfaces of chamber slides, the cells were immobilized on adhesion slides for the staining proceedure. The cells were allowed to attach to adhesion slides, fixed in 1% paraformaldehyde, permeabilized in 0.25% Triton X 100 in PBS, blocked with 1% BSA and stained overnight with 5 µg/ml antibody diluted with 1% BSA in PBS. The secondary antibody was diluted with 1% BSA in PBS according to manufacturers instruction and incubated for 2 hours.

Immunohistochemistry

Supportive validation

Submitted by

Acris Antibodies GmbH (provider)

Main image

Experimental details

Figure 2. Shows K562 (chronic myelogenous leukemia), RDES (Ewings sarcoma), SKES (Ewings) sarcoma and PFSK1 (Primitive neuroectodermal tumor) cells stained with anti-NEUROD1 Antibody Cat.-No AM32710PU-N (Clone BC18-1.11EG) by the indirect immunoperoxidase method. K562 was used as a negative control and showed no staining whereas RDES, SKES-1 and PFSK-1 cells showed weak nuclear but strong cytoplasmic staining. Since the cell lines tested loosely attach to surfaces of chamber slides, the cells were immobilized on adhesion slides for the staining proceedure. The cells were fixed in 1% paraformaldehyde, permeabilized in 0.25% Triton X 100 in PBS, blocked with 1% BSA and stained overnight with 5 µg/ml antibody diluted with 1% BSA in PBS. The peroxidase conjugated anti Mouse Ig secondary antibody was used at 1/1000 in 1% BSA in PBS. DAB substrate was used with nickle chloride enhancement to give dark purple/black staining for positive reaction. The substrate was added for 10 minutes, extensively washed, counterstained in diluted hematoxylene, dehydrated, cleared in xylene and mounted.