Rat astrocytes (DIV 22) cells were fixed with 4% paraformaldehyde and stained for MAP2 (Green) using ab32454 at 1/1000 dilution in ICC/IF analysis.

IHC-P image of ab32454 stained lizard spinal cord. Tissue was fixed in formaldehyde with heat mediated antigen retrieval using citric acid. Tissue was blocked for 10 minutes in 1% B.S.A, and incubated with the antibody (ab32454, 1/250) for 2 hours at 21°C. The secondary antibody used was a goat-anti rabbit IgG conjugated to bitoin (1/250).

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All lanes : Anti-MAP2 antibody - Neuronal Marker (ab32454) at 1 µg/ml

Lane 1 : Mouse Brain (day 0) Tissue Lysate
Lane 2 : Mouse Brain Tissue Lysate
Lane 3 : Rat Brain Tissue Lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

Predicted band size: 199 kDa
Observed band size: 268,280 kDa why is the actual band size different from the predicted?


Exposure time: 2 minutes

Blocking buffer: 2% BSA

Gel type: TA

ab32454 staining MAP2 in mouse brain tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/3000 in blocking buffer) for 2 hours at 21°C in TBS/BSA/azide. An undiluted Biotin-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

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ICC/IF image of ab32454 stained rat PC12 cells. The cells were PFA fixed (10 min), permabilised in PBS-T (20 min) and incubated with the antibody (ab32454, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

All lanes : Anti-MAP2 antibody - Neuronal Marker (ab32454) at 1/1000 dilution

Lanes 1-2 : Trisomic induced Pluripotent Stem Cells (iPSCs)
Lanes 3-4 : Disomic induced Pluripotent Stem Cells (iPSCs)

Predicted band size: 199 kDa

ab32454 staining MAP2 in rat cryopreserved embryonic cortical neurons cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde with picric acid. Samples were incubated with primary antibody (1/2000 in 10mM PBS + 0.3% Triton-X) for 12 hours at 4°C. An Alexa Fluor^® 488-conjugated donkey anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.

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IHC image of MAP2 staining in human cerebral cortex FFPE section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32454, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.