Anti-Calbindin抗体[AF2E5] (ab75524)

概述

  • 产品名称Anti-Calbindin抗体[AF2E5]
    参阅全部 Calbindin 一抗
  • 描述
    小鼠单克隆抗体[AF2E5] to Calbindin
  • 经测试应用适用于: IHC-FoFr, Flow Cyt, ICC/IF, WB, ELISAmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Recombinant full length Human Calbindin protein with a His tag (His-Calbindin) purified from E. coli

  • 阳性对照
    • IMR32 cell lysate.

性能

应用

Our Abpromise guarantee covers the use of ab75524 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
IHC-FoFr 1/2800.
Flow Cyt Use 1µg for 106 cells. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
ICC/IF 1/1000.
WB 1/2000. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa).
ELISA Use at an assay dependent concentration.

靶标

  • 功能Buffers cytosolic calcium. May stimulate a membrane Ca(2+)-ATPase and a 3',5'-cyclic nucleotide phosphodiesterase.
  • 序列相似性Belongs to the calbindin family.
    Contains 5 EF-hand domains.
  • 结构域This protein has four functional calcium-binding sites; potential sites II and VI have lost affinity for calcium.
  • Information by UniProt
  • 数据库链接
  • 别名
    • avian-type antibody
    • CAB27 antibody
    • CALB 1 antibody
    • CALB antibody
    • CALB1 antibody
    • CALB1_HUMAN antibody
    • Calbindin 1 28kDa antibody
    • Calbindin antibody
    • Calbindin D28 antibody
    • D 28K antibody
    • D-28K antibody
    • D28K antibody
    • OTTHUMP00000166027 antibody
    • OTTHUMP00000225441 antibody
    • RTVL H protein antibody
    • Vitamin D dependent calcium binding protein antibody
    • Vitamin D dependent calcium binding protein avian type antibody
    • Vitamin D-dependent calcium-binding protein antibody
    see all

Anti-Calbindin antibody [AF2E5] 图像

  • Anti-Calbindin antibody [AF2E5] (ab75524) at 1/2000 dilution (in blocking buffer for 16 hours at 4°C) + Mouse hippocampus whole tissue lysate at 5 µg

    Secondary
    An HRP-conjugated Horse anti-mouse IgG polyclonal at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 30 kDa
    Observed band size : 25 kDa (why is the actual band size different from the predicted?)


    Exposure time : 15 seconds

    This image is courtesy of an anonymous Abreview

    See Abreview

  • All lanes : Anti-Calbindin antibody [AF2E5] (ab75524) at 1/500 dilution

    Lane 1 : Human kidney tissue lysate - total protein (ab30203)
    Lane 2 : Kidney (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 30 kDa
    Observed band size : 30 kDa


    Exposure time : 90 seconds
  • ab75524 staining rat brain (cortex) sections by IHC-FoFr. The animal was perfused with 4% paraformaldehyde and further post fixed with 4% paraformaldehyde overnight. The tissues were cryoprotected with 30% sucrose and sectioned using a cryostat. Staining with ab75524 at a 1/2800 dilution in PBS-Triton (0.3%) with 0.02% azide was performed for 18h at 25°C. A donkey anti-mouse Alexa488 polyclonal antibody at 1/1000 was used as the secondary antibody.

    See Abreview

  • Overlay histogram showing SH-SY5Y cells stained with ab75524 (red line). The cells were fixed with 80% methanol (5 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 (ab75524, 1µg/1x106 cells) 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 IgG2a [ICIGG2A] (ab91361, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • ICC/IF image of ab75524 stained Hek293 cells. The cells were 4% PFA fixed (10mins) 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 (ab75524, 1:1000 dilution) overnight at +4°C. The secondary antibody (green) was ab96879 Dylight 488 goat anti-mouse IgG (H+L) used at a 1/250 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.

Anti-Calbindin antibody [AF2E5] (ab75524)参考文献

ab75524 has not yet been referenced specifically in any publications.

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Rat Tissue sections (Brain - Cerebral Cortex)
Specification Brain - Cerebral Cortex
Fixative Paraformaldehyde
Permeabilization No
Username

Dr. Sophie Pezet

Verified customer

提交于 Mar 21 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Rat Tissue sections (Brain)
Specification Brain
Fixative Paraformaldehyde
Permeabilization No
Username

Dr. Karine Thibault

Verified customer

提交于 Mar 12 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (hippocampus)
Loading amount 5 µg
Specification hippocampus
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Sep 07 2010

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