Anti-CRM1 antibody

• ICC/IF

Ap17016****

Immunocytochemistry/ Immunofluorescence - Anti-CRM1 antibody (AB24189)

ICC/IF image of ab24189 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab24189 at 1μg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

• IHC-P

Ap16036****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRM1 antibody (AB24189)

IHC image of CRM1 staining in Human normal hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ 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 ab24189, 5μg/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.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• WB

Project1433****

Western blot - Anti-CRM1 antibody (AB24189)

ab24189 detects a band of ~ 100 kDa in HeLa whole and HeLa nuclear cell lysates. This is somewhat smaller than the predicted band size according to Swissprot (123 kDa), however as both these bands are specifically blocked by the addition of the immunizing peptide (ab25749) we believe they represent CRM1.

All lanes:

Western blot - Anti-CRM1 antibody (ab24189) at 1 µg/mL

Lane 1:

HeLa whole cell lysate at 20 µg

Lane 2:

HeLa nuclear lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg with Human CRM1 peptide (<a href='/en-us/products/unavailable/human-crm1-peptide-ab25749'>ab25749</a>)

Lane 4:

HeLa nuclear cell lysate at 20 µg with Human CRM1 peptide (<a href='/en-us/products/unavailable/human-crm1-peptide-ab25749'>ab25749</a>)

Secondary

All lanes:

Goat polyclonal to Rabbit IgG at 1/10000 dilution

Predicted band size: 123 kDa

Observed band size: 100 kDa

false

This image is courtesy of an anonymous Abreview

• WB

Ap17016****

Western blot - Anti-CRM1 antibody (AB24189)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab24189 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-CRM1 antibody (ab24189) at 1 µg/mL

All lanes:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) at 10 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 1/10000 dilution

Predicted band size: 123 kDa

Observed band size: 120 kDa,73 kDa

true

Exposure time: 90s