Key features and details
- Mouse monoclonal [N258/5] to CLC3
- Suitable for: Flow Cyt, WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
- Isotype: IgG1
Product nameAnti-CLC3 antibody [N258/5]
DescriptionMouse monoclonal [N258/5] to CLC3
SpecificityDoes not cross-react with CLCN4 or CLCN5 (based on KO validation results).
Tested applicationsSuitable for: Flow Cyt, WB, IHC-P, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
- WB: Rat brain membrane lysate. IHC-P: FFPE human hippocampus tissue sections. ICC/IF: SK-N-BE cells.
The clone number has been updated from S258-5 to N258/5, both clone numbers name the same antibody clone.
Previously labelled as CLCN3.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Storage instructionsShipped at 4°C. Store at -20ºC.
Storage bufferPreservative: 0.09% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine)
Concentration information loading...
PurityProtein G purified
Our Abpromise guarantee covers the use of ab134285 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|WB||1/1000. Predicted molecular weight: 91 kDa.
1µg/ml of ab134285 is sufficient for detection of CLC3 in 20µg of Rat brain membrane lysate.
FunctionMediates the exchange of chloride ions against protons. Functions as antiporter and contributes to the acidification of the endosome and synaptic vesicle lumen, and may thereby affect vesicle trafficking and exocytosis. May play an important role in neuronal cell function through regulation of membrane excitability by protein kinase C. It could help neuronal cells to establish short-term memory.
Tissue specificityExpressed primarily in tissues derived from neuroectoderm. Within the brain, its expression is particularly evident in the hippocampus, olfactory cortex, and olfactory bulb. Highly expressed in aortic and coronary vascular smooth muscle cells, and aortic endothelial cells. Also expressed in tracheal and alveolar epithelial cells, and intima and media of the pulmonary vessels. Expressed in bronchus and colon (at protein level).
Sequence similaritiesBelongs to the chloride channel (TC 2.A.49) family. ClC-3/CLCN3 subfamily.
Contains 2 CBS domains.
DomainIsoform 2 contains a C-terminal PDZ-binding motif mediating the interaction with GOPC.
Cellular localizationMembrane. Early endosome membrane. Late endosome membrane. Cytoplasmic vesicle > secretory vesicle membrane. Isoform 1 is localized mainly in early and late endosomes and Membrane. Early endosome membrane. Late endosome membrane. Golgi apparatus membrane. Isoform 2 partially colocalized with isoform 1 but is mainly enriched in the Golgi.
- Information by UniProt
- Chloride channel 3 antibody
- Chloride channel protein 3 antibody
- Chloride channel, voltage-sensitive 3 antibody
Anti-CLC3 antibody [N258/5] (ab134285) at 1/200 dilution + Rat brain membrane lysate at 15 µg
Goat Anti-Mouse IgG: HRP at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 91 kDa
4% formaldehyde-fixed SK-N-BE cells stained for CLC3 (green) using ab134285 at 1/100 dilution in ICC/IF.
Secondary Antibody: Goat Anti-Mouse ATTO 488 at 1/100 for 60 minutes at room temperature. Counterstain: Phalloidin Texas Red F-Actin stain; DAPI (blue) nuclear stain at 1/1000, 1/5000 for 60 minutes room temperature, 5 minutes room temperature.
(A) DAPI (blue) nuclear stain. (B) Phalloidin Texas Red F-Actin stain. (C) ab134285. (D) Composite.
IHC image of CLC3 staining in human 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 ab134285, 10µg/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.
Overlay histogram showing SH-SY5Y cells stained with ab134285 (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 (ab134285, 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.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab134285 has been referenced in 3 publications.
- Cheng W et al. Chloride channel 3 (CIC-3) predicts the tumor size in hepatocarcinoma. Acta Histochem 121:284-288 (2019). PubMed: 30678806
- Xu ZZ et al. 17ß-Estradiol Attenuates Neuropathic Pain Caused by Spared Nerve Injury by Upregulating CIC-3 in the Dorsal Root Ganglion of Ovariectomized Rats. Front Neurosci 13:1205 (2019). PubMed: 31787875
- Deng Z et al. Estradiol activates chloride channels via estrogen receptor-a in the cell membranes of osteoblasts. Am J Physiol Cell Physiol 313:C162-C172 (2017). PubMed: 28468943