Anti-CLC3 antibody [N258/5] (ab134285)
Key features and details
- Mouse monoclonal [N258/5] to CLC3
- Suitable for: Flow Cyt, WB, IHC-P, ICC/IF
- Reacts with: Rat, Human
- Isotype: IgG1
Overview
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Product name
Anti-CLC3 antibody [N258/5]
See all CLC3 primary antibodies -
Description
Mouse monoclonal [N258/5] to CLC3 -
Host species
Mouse -
Specificity
Does not cross-react with CLCN4 or CLCN5 (based on KO validation results). -
Tested applications
Suitable for: Flow Cyt, WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Rat, Human -
Immunogen
Synthetic peptide corresponding to Rat CLC3 aa 98-115 (N terminal).
Sequence:CKDRERHRRINSKKKESA
Database link: NP_445815.2 -
Positive control
- WB: Rat brain membrane lysate. IHC-P: FFPE human hippocampus tissue sections. ICC/IF: SK-N-BE cells.
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General notes
The clone number has been updated from S258-5 to N258/5, both clone numbers name the same antibody clone.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20ºC. -
Storage buffer
Preservative: 0.09% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
N258/5 -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
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Flow Cyt |
Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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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. |
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IHC-P |
1/1000.
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ICC/IF |
1/100.
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Notes |
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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. |
IHC-P
1/1000. |
ICC/IF
1/100. |
Target
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Function
Mediates 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 specificity
Expressed 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 similarities
Belongs to the chloride channel (TC 2.A.49) family. ClC-3/CLCN3 subfamily.
Contains 2 CBS domains. -
Domain
Isoform 2 contains a C-terminal PDZ-binding motif mediating the interaction with GOPC. -
Post-translational
modificationsN-glycosylated. -
Cellular localization
Membrane. 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
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Database links
- Entrez Gene: 1182 Human
- Entrez Gene: 84360 Rat
- Omim: 600580 Human
- SwissProt: P51790 Human
- SwissProt: P51792 Rat
- Unigene: 481186 Human
- Unigene: 4175 Rat
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Alternative names
- Chloride channel 3 antibody
- Chloride channel protein 3 antibody
- Chloride channel, voltage-sensitive 3 antibody
see all
Images
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Anti-CLC3 antibody [N258/5] (ab134285) at 1/200 dilution + Rat brain membrane lysate at 15 µg
Secondary
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.
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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.
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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.
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
References (6)
ab134285 has been referenced in 6 publications.
- Lei W et al. Disulfiram-copper activates chloride currents and induces apoptosis with tyrosine kinase in prostate cancer cells. Asia Pac J Clin Oncol 18:e46-e55 (2022). PubMed: 33608991
- Zhang B et al. ClC-3 induction protects against cerebral ischemia/reperfusion injury through promoting Beclin1/Vps34-mediated autophagy. Hum Cell 33:1046-1055 (2020). PubMed: 32772229
- 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
- Wang D et al. ClC-3 Promotes Osteogenic Differentiation in MC3T3-E1 Cell After Dynamic Compression. J Cell Biochem 118:1606-1613 (2017). PubMed: 27922190