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Signal Transduction Signaling Pathway Calcium Signaling Calcium Binding Proteins

Anti-Calsequestrin 2 + Calsequestrin 1抗体(ab3516)

  • Datasheet
Reviews (4)Q&A (2)References (59)

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Western blot - Anti-Calsequestrin 2 + Calsequestrin 1 antibody (ab3516)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calsequestrin 2 + Calsequestrin 1 antibody (ab3516)
  • Immunocytochemistry/ Immunofluorescence - Anti-Calsequestrin 2 + Calsequestrin 1 antibody (ab3516)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calsequestrin 2 + Calsequestrin 1 antibody (ab3516)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calsequestrin 2 + Calsequestrin 1 antibody (ab3516)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calsequestrin 2 + Calsequestrin 1 antibody (ab3516)
  • Immunocytochemistry/ Immunofluorescence - Anti-Calsequestrin 2 + Calsequestrin 1 antibody (ab3516)

Key features and details

  • Rabbit polyclonal to Calsequestrin 2 + Calsequestrin 1
  • Suitable for: ICC/IF, WB, IP, IHC-P
  • Reacts with: Mouse, Rat, Sheep, Rabbit, Dog, Human, Pig, Rainbow trout
  • Isotype: IgG

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概述

  • 产品名称

    Anti-Calsequestrin 2 + Calsequestrin 1抗体
  • 描述

    兔多克隆抗体to Calsequestrin 2 + Calsequestrin 1
  • 宿主

    Rabbit
  • 特异性

    This antibody recognizes both cardiac and skeletal muscle calsequestrin.
  • 经测试应用

    适用于: ICC/IF, WB, IP, IHC-Pmore details
  • 种属反应性

    与反应: Mouse, Rat, Sheep, Rabbit, Dog, Human, Pig, Rainbow trout
  • 免疫原

    Full length native protein (purified) corresponding to Dog Calsequestrin 2. The immunogen is purified canine cardiac calsequestrin.

  • 常规说明

    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

性能

  • 形式

    Liquid
  • 存放说明

    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.
  • 存储溶液

    Preservative: 0.05% Sodium azide
  • Concentration information loading...
  • 纯度

    Whole antiserum
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Signal Transduction
    • Signaling Pathway
    • Calcium Signaling
    • Calcium Binding Proteins
    • Signal Transduction
    • Protein Trafficking
    • ER Proteins
    • Cardiovascular
    • Heart
    • Cardiac metabolism
    • Cardiovascular
    • Heart
    • Cardiac arrhythmias
    • Metabolism
    • Types of disease
    • Cancer

相关产品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

应用

The Abpromise guarantee

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.
说明
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.

IP
Use at an assay dependent concentration.
IHC-P
1/100 - 1/500.

靶标

  • 细胞定位

    Calsequestrin 2: Sarcoplasmic reticulum lumen. This isoform of calsequestrin occurs in the sarcoplasmic reticulum's terminal cisternae luminal spaces of cardiac and slow skeletal muscle cells. Calsequestrin 1: 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.
  • 数据库链接

    • Entrez Gene: 483134 Dog
    • Entrez Gene: 844 Human
    • Entrez Gene: 845 Human
    • Entrez Gene: 12372 Mouse
    • Entrez Gene: 12373 Mouse
    • Entrez Gene: 100009095 Rabbit
    • Entrez Gene: 29209 Rat
    • Entrez Gene: 686019 Rat
    • Omim: 114250 Human
    • Omim: 114251 Human
    • SwissProt: P12637 Dog
    • SwissProt: O14958 Human
    • SwissProt: P31415 Human
    • SwissProt: O09161 Mouse
    • SwissProt: O09165 Mouse
    • SwissProt: P07221 Rabbit
    • SwissProt: P19633 Rat
    • SwissProt: P51868 Rat
    • Unigene: 57975 Human
    • Unigene: 632476 Human
    • Unigene: 15343 Mouse
    • Unigene: 10111 Rat
    see all

图片

  • Western blot - Anti-Calsequestrin 2 + Calsequestrin 1 antibody (ab3516)
    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 kDa why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calsequestrin 2 + Calsequestrin 1 antibody (ab3516)
    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)
    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)
    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)
    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)
    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)
    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.

    See Abreview

实验方案

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

数据表及文件

  • Datasheet download

    Download

文献 (59)

发表研究结果有使用 ab3516?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab3516 被引用在 59 文献中.

  • Wyckelsma VL  et al. Loss of a-actinin-3 during human evolution provides superior cold resilience and muscle heat generation. Am J Hum Genet 108:446-457 (2021). PubMed: 33600773
  • Monteiro DA  et al. Fluid shear stress generates a unique signaling response by activating multiple TGFß family type I receptors in osteocytes. FASEB J 35:e21263 (2021). PubMed: 33570811
  • Mia S  et al. Impact of obesity on day-night differences in cardiac metabolism. FASEB J 35:e21298 (2021). PubMed: 33660366
  • Shankar TS  et al. Cardiac-specific deletion of voltage dependent anion channel 2 leads to dilated cardiomyopathy by altering calcium homeostasis. Nat Commun 12:4583 (2021). PubMed: 34321484
  • Oldfield CJ  et al. Muscle-specific sirtuin 3 overexpression does not attenuate the pathological effects of high-fat/high-sucrose feeding but does enhance cardiac SERCA2a activity. Physiol Rep 9:e14961 (2021). PubMed: 34405591
View all Publications for this product

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1-6 of 6 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Sample
Mouse Cell (myocyte)
Specification
myocyte
Permeabilization
No
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Jun 14 2013

Western blot

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
5 µg
Gel Running Conditions
Reduced Denaturing (8)
Sample
Human Tissue lysate - whole (Heart)
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Jun 13 2013

Western blot abreview for Anti-Calsequestrin antibody

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Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Pig Tissue lysate - whole (heart)
Loading amount
7.5 µg
Specification
heart
Gel Running Conditions
Reduced Denaturing (7.5%)
Blocking step
Invitrogen 00-0105 membrane blocking agent as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Apr 05 2013

Western blot abreview for Anti-Calsequestrin antibody

Average
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (heart, ventricular tissue)
Loading amount
60 µg
Specification
heart, ventricular tissue
Gel Running Conditions
Reduced Denaturing (4~12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Feb 02 2008

Question

Can you tell me which abcam product best fits the inquiry attached?  I think it is ab3516 I also found ab97474 and ab32801.

Read More

Abcam community

Verified customer

Asked on Nov 04 2011

Answer

I would recommened ab3516, it has been tested in more applications and species.

Read More

Abcam Scientific Support

回复于 Nov 04 2011

Question

Wondering if you could inform us about the target sequence for the rabbit polyclonal to Calsequestrin (ab3516)antibody.

Read More

Abcam community

Verified customer

Asked on Jun 25 2007

Answer

Thank you for your enquiry. This anti-Calsequestrin antibody (ab3516) was created against purified canine cardiac calsequestrin. Unfortunately, the exact epitope sequence has not been mapped at this time. I am sorry I can't be of more assistance in this occasion but should you require any further information, please do not hesitate to contact me.

Read More

Abcam Scientific Support

回复于 Jun 25 2007

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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