Product nameAnti-WIPI2 antibody [2A2]
See all WIPI2 primary antibodies
DescriptionMouse monoclonal [2A2] to WIPI2
Tested applicationsSuitable for: ICC/IF, IP, IHC-P, IHC-Fr, WB, Flow Cytmore details
Species reactivityReacts with: Mouse, Human
Synthetic peptide corresponding to Human WIPI2 aa 400-500 (C terminal).
- This antibody gave a positive signal in Human Skeletal Muscle, Human Placenta, Mouse Placenta, Mouse Testis as well as the following whole cell lysates: HeLa, and NIH3T3. IF/ICC: HeLa cells (50mM chloroquine for 24h) Flow Cyt: HeLa cells IHC-P: Human skeletal muscle (normal)
This antibody clone is manufactured by Abcam. We welcome customer feedback and would appreciate any comments regarding this product and the data presented.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferPreservative: 0.02% Sodium azide
Concentration information loading...
PurityProtein A purified
Light chain typekappa
Our Abpromise guarantee covers the use of ab105459 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration. Fix with Acetone.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 49 kDa.|
|Flow Cyt||Use 0.1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
FunctionProbable early component of the autophagy machinery being involved in formation of preautophagosomal structures and their maturation into mature phagosomes in response to PtdIns3P. May bind PtdIns3P.
Tissue specificityUbiquitously expressed (at protein level). Highly expressed in heart, skeletal muscle and pancreas. Expression is down-regulated in pancreatic and in kidney tumors.
Sequence similaritiesBelongs to the WD repeat SVP1 family.
Contains 3 WD repeats.
Cellular localizationPreautophagosomal structure membrane. Enriched at preautophagosomal structure membranes in response to ptdIns3P.
- Information by UniProt
- ATG18B antibody
- Atg21 antibody
- CGI 50 antibody
All lanes : Anti-WIPI2 antibody [2A2] (ab105459) at 1 µg/ml
Lane 1 : Human skeletal muscle tissue lysate
Lane 2 : Human placenta tissue lysate
Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 4 : NIH 3T3 (Mouse) Whole Cell Lysate
Lane 5 : Pancreas (Mouse) Tissue Lysate
Lane 6 : Testis (Mouse) Tissue Lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) (ab65485) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 49 kDa
Exposure time: 1 minute
Abcam recommends using milk as the blocking agent.
IHC image of WIPI2 staining in a section of formalin-fixed paraffin-embedded normal human skeletal muscle* performed on a Leica BONDTM 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 20mins. The section was then incubated with ab105459, 1ug/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.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
All lanes : Anti-WIPI2 antibody [2A2] (ab105459) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Wipi2 knockout HAP1 whole cell lysate
Lane 3 : Saos2 whole cell lysate
Lane 4 : CACO2 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 49 kDa
Observed band size: 49 kDa
Lanes 1 - 4: Merged signal (red and green). Green - ab105459 observed at 49 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab105459 was shown to recognize WIPI2 in wild-type HAP1 cells as signal was lost at the expected MW in Wipi2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Wipi2 knockout samples were subjected to SDS-PAGE. Ab105459 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
ab105459 staining WIPI2 in HeLa cells. The cells were treated with 50uM chloroquine for 24h and then fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab105459 at 1ugml then detected with an Alexa Fluor® 488 goat anti-rabbit secondary antibody (ab150081) at a 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue), and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at a 1/250 dilution (shown in red).
Overlay histogram showing HeLa cells stained with ab105459 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab105459, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
This product has been referenced in:
- Padman BS et al. LC3/GABARAPs drive ubiquitin-independent recruitment of Optineurin and NDP52 to amplify mitophagy. Nat Commun 10:408 (2019). Read more (PubMed: 30679426) »
- Liu Y et al. Niclosamide Triggers Non-Canonical LC3 Lipidation. Cells 8:N/A (2019). Read more (PubMed: 30875964) »