Anti-Calsequestrin 2 + Calsequestrin 1抗体
兔多克隆抗体to Calsequestrin 2 + Calsequestrin 1
Rabbit
This antibody recognizes both cardiac and skeletal muscle calsequestrin.
适用于: ICC/IF, WB, IP, IHC-Pmore details
与反应: Mouse, Rat, Sheep, Rabbit, Dog, Human, Pig, Rainbow trout
Full length native protein (purified) corresponding to Dog Calsequestrin 2. The immunogen is purified canine cardiac calsequestrin.
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Liquid
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.
Preservative: 0.05% Sodium azide
Whole antiserum
多克隆
IgG
Abpromise™承诺保证使用ab3516于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| ICC/IF | (1) | Use at an assay dependent concentration. |
| WB | (3) | 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. |
| IP | Use at an assay dependent concentration. | |
| IHC-P | 1/100 - 1/500. |
Entrez Gene: 844 Human
Entrez Gene: 845 Human
Entrez Gene: 12372 Mouse
Entrez Gene: 12373 Mouse
Entrez Gene: 100009095 Rabbit
Omim: 114250 Human
Omim: 114251 Human
SwissProt: O14958 Human
SwissProt: P31415 Human
SwissProt: O09161 Mouse
SwissProt: O09165 Mouse
SwissProt: P07221 Rabbit
Unigene: 57975 Human
Unigene: 632476 Human
Unigene: 15343 Mouse
Unigene: 10111 Rat
Western blot - Anti-Calsequestrin 2 + Calsequestrin 1 antibody (ab3516)
All lanes : Anti-Calsequestrin 2 + Calsequestrin 1 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 kDawhy is the actual band size different from the predicted?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calsequestrin 2 + Calsequestrin 1 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.
Immunocytochemistry/ Immunofluorescence - Anti-Calsequestrin 2 + Calsequestrin 1 antibody (ab3516)
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calsequestrin 2 + Calsequestrin 1 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calsequestrin 2 + Calsequestrin 1 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calsequestrin 2 + Calsequestrin 1 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 2 + Calsequestrin 1 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.
