Anti-Calsequestrin 抗体 (ab3516)
Key features and details
- Rabbit polyclonal to Calsequestrin
- Suitable for: ICC/IF, WB, IHC-P
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
製品の概要
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製品名
Anti-Calsequestrin antibody -
製品の詳細
Rabbit polyclonal to Calsequestrin -
由来種
Rabbit -
特異性
This antibody recognizes both cardiac and skeletal muscle calsequestrin. -
アプリケーション
適用あり: ICC/IF, WB, IHC-Pmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Other Immunogen Type corresponding to Dog Calsequestrin. The immunogen is purified canine cardiac calsequestrin.
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特記事項
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.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
バッファー
Preservative: 0.05% Sodium azide -
Concentration information loading...
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精製度
Immunogen affinity purified -
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
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Compatible Secondaries
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Isotype control
アプリケーション
Our Abpromise guarantee covers the use of ab3516 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 |
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ICC/IF | Use at an assay dependent concentration. | |
WB | 1/1000 - 1/10000. This antibody detects an ~55 kDa protein representing Calsequestrin from canine cardiac extract. Additional bands at 97 kDa may be observed and have been reported to be Calsequestrin-like proteins. |
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IHC-P | 1/100 - 1/500. |
ターゲット情報
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機能
Calsequestrin is a high-capacity, moderate affinity, calcium-binding protein and thus acts as an internal calcium store in muscle. The release of calcium bound to calsequestrin through a calcium release channel triggers muscle contraction. Binds 40 to 50 moles of calcium. Also binds laminin. -
配列類似性
Belongs to the calsequestrin family. -
細胞内局在
Sarcoplasmic reticulum lumen. Mitochondrion. This isoform of calsequestrin occurs in the sarcoplasmic reticulum's terminal cisternae luminal spaces of fast skeletal muscle cells. Also mitochondrial according to PubMed:7945294. - Information by UniProt
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参照データベース
- Entrez Gene: 844 Human
- Entrez Gene: 845 Human
- Entrez Gene: 12372 Mouse
- Entrez Gene: 12373 Mouse
- Entrez Gene: 29209 Rat
- Omim: 114250 Human
- Omim: 114251 Human
- SwissProt: O14958 Human
see all -
別名
- Calmitin antibody
- Calsequestrin 1 antibody
- Calsequestrin 1 fast twitch skeletal muscle antibody
see all
画像
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All lanes : Anti-Calsequestrin antibody (ab3516) at 1/5000 dilution
Lane 1 : RD (Human muscle rhabdomyosarcoma cell line) whole cell lysate
Lane 2 : L6 (Rat skeletal muscle cell line) whole cell lysate
Lane 3 : Mouse heart tissue lysate
Lysates/proteins at 25 µg per lane.
Secondary
All lanes : HRP-conjugated secondary antibody
Observed band size: 55 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calsequestrin antibody (ab3516)
ab3516 labelling Calsequestrin in the cytoplasm of human skeletal muscle tissue (right) compared with a negative control (left) by Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS. Tissue sections were incubated with primary antibody (1:200 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-rabbit was used as the secondary antibody, followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
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Immunofluorescent analysis of Calsequestrin (green) showing staining in the cytoplasm and nucleus of C2C12 (Mouse myoblast cell line) cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with ab3516 in 3% BSA-PBS at a dilution of 1/100 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calsequestrin antibody (ab3516)
ab3516 labelling Calsequestrin in the cytoplasm of mouse heart tissue (right) compared with a negative control (left) by Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS. Tissue sections were incubated with primary antibody (1:200 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-rabbit was used as the secondary antibody, followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calsequestrin antibody (ab3516)
ab3516 labelling Calsequestrin in the cytoplasm of human heart tissue (right) compared with a negative control (left) by Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS. Tissue sections were incubated with primary antibody (1:200 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-rabbit was used as the secondary antibody, followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calsequestrin antibody (ab3516)
ab3516 (1µg/ml) staining Calsequestrin in human skeletal muscle using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of discrete organelles within the cytoplasm.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required. -
Immunocytochemistry/ Immunofluorescence - Anti-Calsequestrin antibody (ab3516)This image is courtesy of an anonymous Abreview
ab3516 staining Calsequestrin in mouse myocyte cell by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde and blocked with 10% serum for 1 hour at 25°C. Samples were incubated with primary antibody (1/50 in serum) for 18 hours at 4°C. An Alexa Fluor®488-conjugated Goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.
プロトコール
データシートおよび資料
参考文献 (47)
ab3516 は 47 報の論文で使用されています。
- Eckhardt J et al. Molecular basis of impaired extraocular muscle function in a mouse model of congenital myopathy due to compound heterozygous Ryr1 mutations. Hum Mol Genet 29:1330-1339 (2020). PubMed: 32242214
- Asghari P et al. Cardiac ryanodine receptor distribution is dynamic and changed by auxiliary proteins and post-translational modification. Elife 9:N/A (2020). PubMed: 31916935
- Michailowsky V et al. Defects in sarcolemma repair and skeletal muscle function after injury in a mouse model of Niemann-Pick type A/B disease. Skelet Muscle 9:1 (2019). PubMed: 30611303
- Christiansen D et al. A fast, reliable and sample-sparing method to identify fibre types of single muscle fibres. Sci Rep 9:6473 (2019). PubMed: 31019216
- Wang Q et al. Two pools of IRE1a in cardiac and skeletal muscle cells. FASEB J N/A:fj201802626R (2019). PubMed: 31051095