Anti-beta 1 Spectrin抗体[4C3] (ab2808)

概述

  • 产品名称Anti-beta 1 Spectrin抗体[4C3]
    参阅全部 beta 1 Spectrin 一抗
  • 描述
    小鼠单克隆抗体[4C3] to beta 1 Spectrin
  • 特异性Detects spectrin from erythrocytes, brain and muscle cells. This antibody has been shown to specifically detect the two known alternatively spliced forms of spectrin, beta-1 epsilon-1, present in erythrocytes, and beta-1 epsilon-2, present in nerve and striated muscle cells. It does not cross-react with alpha-2 spectrin or either of the fodrin subunits.
  • 经测试应用适用于: Flow Cyt, IHC-P, ICC/IF, WB, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Full length native protein (purified) corresponding to Human beta 1 Spectrin. Purified human erythrocyte beta-1 spectrin.

性能

应用

Our Abpromise guarantee covers the use of ab2808 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
Flow Cyt 1/50.

IHC-P 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/10 - 1/200.
WB 1/100.
IHC-P 1/20 - 1/200.

Perform antigen retrieval method using 10mM sodium citrate (pH 6.0) and microwaved for 8-15 min before commencing with IHC staining protocol.

靶标

  • 功能Spectrin is the major constituent of the cytoskeletal network underlying the erythrocyte plasma membrane. It associates with band 4.1 and actin to form the cytoskeletal superstructure of the erythrocyte plasma membrane.
  • 疾病相关Defects in SPTB are the cause of elliptocytosis type 3 (EL3) [MIM:182870]. EL3 is a Rhesus-unlinked form of hereditary elliptocytosis, a genetically heterogeneous, autosomal dominant hematologic disorder. It is characterized by variable hemolytic anemia and elliptical or oval red cell shape.
    Defects in SPTB are the cause of spherocytosis type 2 (SPH2) [MIM:182870]; also known as hereditary spherocytosis type 2 (HS2). Spherocytosis is a hematologic disorder leading to chronic hemolytic anemia and characterized by numerous abnormally shaped erythrocytes which are generally spheroidal. SPH2 is characterized by severe hemolytic anemia. Inheritance is autosomal dominant.
  • 序列相似性Belongs to the spectrin family.
    Contains 2 CH (calponin-homology) domains.
    Contains 17 spectrin repeats.
  • 翻译后修饰The first phosphorylation event occurs on Ser-2114, followed by Ser-2125, Ser-2123, Ser-2128, Ser-2117, and Thr-2110.
  • 细胞定位Cytoplasm > cytoskeleton. Cytoplasm > cell cortex.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Beta I spectrin antibody
    • Beta spectrin antibody
    • Beta-I spectrin antibody
    • EL3 antibody
    • erythrocyte antibody
    • HS2 antibody
    • HSpTB1 antibody
    • Membrane cytoskeletal protein antibody
    • Spectrin beta antibody
    • Spectrin beta chain antibody
    • Spectrin beta chain erythrocyte antibody
    • Spectrin beta erythrocytic (includes spherocytosis clinical type I) antibody
    • Spectrin beta erythrocytic antibody
    • SPH2 antibody
    • sptB antibody
    • SPTB1 antibody
    • SPTB1_HUMAN antibody
    see all

Anti-beta 1 Spectrin antibody [4C3] 图像

  • Immunocytochemistry/Immunofluorescence analysis of beta 1 Spectrin (green) showing staining in the cytoplasm of A431 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 incubated with ab2808 in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4 ºC 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.

  • Immunocytochemistry/Immunofluorescence analysis of beta 1 Spectrin (green) showing staining in the cytoplasm of Hela 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 incubated with ab2808 in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 ºC 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.

  • Anti-beta 1 Spectrin antibody [4C3] (ab2808) at 1/1000 dilution + HeLa cell lysate at 25 µg
  • ab2808 labelling beta 1 Spectrin in the cytoplasm of Human prostate carcinoma (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. Tissue sections were incubated with the primary antibody (1:20 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-mouse 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.

  • ab2808 labelling beta 1 Spectrin in the membrane of Human skeletal muscle (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. Tissue sections were incubated with the primary antibody (1:200 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-mouse 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.

  • ab2808 labelling beta 1 Spectrin in the cytoplasm and membrane of Mouse skeletal muscle (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. Tissue sections were incubated with the primary antibody (1:20 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-mouse 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.

  • IHC image of ab2808 staining in human skeletal muscle formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab2808, 1/200 dilution, 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.

  • Flow cytometry analysis of beta 1 Spectrin showing positive staining in the cytoplasm of SH-SY5Y cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant, adding 90% methanol and incubated for 10 minutes at room temperature. Follwing penetration, cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2808 (1 ug/test) for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

  • Flow cytometry analysis of beta 1 Spectrin showing positive staining in the cytoplasm of NIH/3T3 cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant, adding 90% methanol and incubated for 10 minutes at room temperature. Follwing penetration, cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2808 (0.5 ug/test) for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

  • Flow cytometry analysis of beta 1 Spectrin showing positive staining in the cytoplasm of Hela cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant, adding 90% methanol and incubated for 10 minutes at room temperature. Follwing penetration, cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2808 (1 ug/test) for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

  • Overlay histogram showing SH-SY5Y cells stained with ab2808 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab2808, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

Anti-beta 1 Spectrin antibody [4C3] (ab2808)参考文献

This product has been referenced in:
  • Martin PT  et al. A comparative study of N-glycolylneuraminic acid (Neu5Gc) and cytotoxic T cell (CT) carbohydrate expression in normal and dystrophin-deficient dog and human skeletal muscle. PLoS One 9:e88226 (2014). ICC/IF ; Human . Read more (PubMed: 24505439) »
  • Chan MM  et al. Hematopoietic protein-1 regulates the actin membrane skeleton and membrane stability in murine erythrocytes. PLoS One 8:e54902 (2013). WB, ICC/IF ; Mouse . Read more (PubMed: 23424621) »

See all 6 Publications for this product

Product Wall

Thank you for your phone call. I have enquired with the originators of these two antibodies and for ab2808, rat skeletal muscle homogenate should be used as a positive control and for ab2495, the originator used myc-tagged human PDK-1 virus infected Sf...

Read More

It has not been tested if ab2808 detects beta-1 in cardiac muscle but the the originator thinks that it may.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"