Anti-CaMKII alpha (phospho T286) 抗体 (ab5683)
製品の概要
-
製品名Anti-CaMKII alpha (phospho T286) antibody
CaMKII alpha 一次抗体 製品一覧 -
製品の詳細Rabbit polyclonal to CaMKII alpha (phospho T286)
-
特異性It does cross-react with CAMKII beta protein
-
アプリケーション適用あり: IHC-Fr, ICC/IF, WB, IHC (PFA fixed)more details
-
種交差性交差種: Mouse, Rat, Rabbit, Human
-
免疫原
Synthetic phosphopeptide derived from a region of human CaM Kinase II alpha that contains threonine 286.
-
ポジティブ･コントロール
- Rat brain lysate.
-
特記事項
Calcium/calmodulin-dependent protein kinase II alpha (CaM Kinase II alpha) is a 50 kDa member of CaM Kinase II family of serine-threonine kinases that transduce Ca2+ signals to several target proteins, including ion channels and transcription activators. CaM Kinase II is predominantly expressed in two isoforms in the brain: alpha and beta. CaM Kinase II plays an important role in neuronal plasticity and memory formation, and exerts both calcium-calmodulin-dependent and -independent activities. Autophosphorylation of CaM Kinase II alpha on threonine 286 allows the kinase to switch from a calmodulin-dependent to a calmodulin-independent state, and is required for various cellular functions including hippocampal long-term potentiation (LTP), special learning, and hippocampus-dependent memory.
製品の特性
-
製品の状態Liquid
-
保存方法Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
-
バッファーPreservative: 0.05% Sodium Azide
Constituents: 50% Glycerol, PBS, 1mg/ml BSA. pH 7.3 -
Concentration information loading... -
精製度Immunogen affinity purified
-
特記事項（精製）The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated CaM Kinase II alpha. The final product is generated by affinity chromatography using a CaM Kinase II alpha-derived peptide that is phosphorylated at threonine 286.
-
一次抗体 備考Calcium/calmodulin-dependent protein kinase II alpha (CaM Kinase II alpha) is a 50 kDa member of CaM Kinase II family of serine-threonine kinases that transduce Ca2+ signals to several target proteins, including ion channels and transcription activators. CaM Kinase II is predominantly expressed in two isoforms in the brain: alpha and beta. CaM Kinase II plays an important role in neuronal plasticity and memory formation, and exerts both calcium-calmodulin-dependent and -independent activities. Autophosphorylation of CaM Kinase II alpha on threonine 286 allows the kinase to switch from a calmodulin-dependent to a calmodulin-independent state, and is required for various cellular functions including hippocampal long-term potentiation (LTP), special learning, and hippocampus-dependent memory.
-
ポリ/モノポリクローナル
-
アイソタイプIgG
-
研究分野
関連製品
-
Compatible Secondaries
-
Immunohistochemistry kits
アプリケーション
Our Abpromise guarantee covers the use of ab5683 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| アプリケーション | Abreviews | 特記事項 |
|---|---|---|
| IHC-Fr | Use at an assay dependent concentration. | |
| ICC/IF | Use a concentration of 5 µg/ml. | |
| WB | 1/1000. Detects a band of approximately 50 kDa. | |
| IHC (PFA fixed) | 1/300. |
ターゲット情報
-
機能CaM-kinase II (CAMK2) is a prominent kinase in the central nervous system that may function in long-term potentiation and neurotransmitter release. Member of the NMDAR signaling complex in excitatory synapses it may regulate NMDAR-dependent potentiation of the AMPAR and synaptic plasticity.
-
配列類似性Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CaMK subfamily.
Contains 1 protein kinase domain. -
細胞内局在Cell junction > synapse > presynaptic cell membrane. Cell junction > synapse. Postsynaptic lipid rafts.
- Information by UniProt
-
参照データベース
- Entrez Gene: 815 Human
- Entrez Gene: 12322 Mouse
- Entrez Gene: 25400 Rat
- Omim: 114078 Human
- SwissProt: Q9UQM7 Human
- SwissProt: P11798 Mouse
- SwissProt: P11275 Rat
- Unigene: 725992 Human
see all -
別名
- Alpha CaMKII antibody
- Calcium calmodulin dependent protein kinase II antibody
- Calcium/calmodulin dependent protein kinase II alpha B subunit antibody
see all
Anti-CaMKII alpha (phospho T286) antibody 画像
-
Immunohistochemistry (PFA fixed) - Anti-CaMKII alpha (phospho T286) antibody (ab5683)This image is courtesy of Sophie Pezet, Univ London Kings Coll, United KingdomImmunofluorescent staining for CaMKII alpha phospho (T286) using ab5683 (1/300, incubated for 18 hours) in rat spinal cord. To induce CaMKII alpha phospho (T286) protein expression, a noxious stimulus was administered to the rat 5 minutes prior to 4% PFA perfusion fixation (this is a known paradigm for inducing phosphorylation CaMKII in some spinal neurons). The resulting immunofluorescent staining for CaMKII alpha phospho (T286) protein is observed in the cytoplasm of many dorsal horn spinal neurons (surrounding the central canal [A] or in the ventral horn [B]). Omition of primary antibody resulted in a lack of staining (data not shown).
Protocol details: Tissue was prepared by 4% paraformaldehyde cardiac perfusion fixation. Tissue was frozen on dry ice and then embedded in OCT compound and cut on cryostat. An antigen retrieval step was not neccesary for the IHC protocol. -
Immunocytochemistry/ Immunofluorescence - Anti-CaMKII alpha (phospho T286) antibody (ab5683)ICC/IF image of ab5683 stained PC12 cells. The cells were 4% formaldehyde fixed (10 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 (ab5683, 5µg/ml) overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. -
Western blot - Anti-CaMKII alpha (phospho T286) antibody (ab5683)Peptide Competition and Phosphatase Treatment: Rat brain lysates were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-4) or treated with lambda (ë) phosphatase (5) and blocked with a 5% BSA-TBST buffer for one hour at room temperature, then incubated with ab5683 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 5), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphothreonine-containing peptide (3), or, the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignal method. The data show that only the peptide corresponding to CaM Kinase II alpha [pT286] blocks the antibody signal. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific. Peptide Competiti -
Western blot - Anti-CaMKII alpha (phospho T286) antibody (ab5683)This image is courtesy of an Abreview submitted by Dr. Byung-il ChoiThis image is courtesy of an Abreview submitted by Dr. Byung-il Choi
Primary Antibody 1/1000 24 hours at 4°C
Secondary Antibody (1/5000): HRP-conjugated Goat polyclonal to rabbit IgG
Blocking step: 5% BSA for 12 hours at 4°C
プロトコール
Anti-CaMKII alpha (phospho T286) antibody (ab5683) 使用論文
This product has been referenced in:
- Kugathasan P et al. In vivo and in vitro effects of vortioxetine on molecules associated with neuroplasticity. J Psychopharmacol 31:365-376 (2017). WB . Read more (PubMed: 27678087) »
- Haas LT et al. Metabotropic glutamate receptor 5 couples cellular prion protein to intracellular signalling in Alzheimer's disease. Brain 139:526-46 (2016). WB, ICC/IF ; Mouse . Read more (PubMed: 26667279) »
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"