製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to Calbindin
  • アプリケーション
    適用あり: IP, WB, IHC-P, IHC-Fr, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Human
    交差が予測される動物種: Rat, Rabbit, Horse, Chicken, Hamster, Cow
  • 免疫原

    Synthetic peptide:

    MCGKEFNKAFELYDQ

    , corresponding to amino acids 185-199 of Human Calbindin

  • ポジティブ・コントロール
    • Rat brain

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab25085 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
IP Use a concentration of 5 µg/ml.
WB Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 28 kDa.
IHC-P 1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 - 5 µg/ml.

ターゲット情報

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  • Calbindin was immunoprecipitated using 0.5mg Rat Brain tissue extract, 5µg of Rabbit polyclonal to Calbindin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, Rat Brain tissue extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab25085.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).<

    Band: 28kDa; Calbindin

  • ab25085 staining human Calbindin D28 in Human cerebellum by immunohistochemistry formalin fixed, paraffin embedded.
  • ICC/IF image of ab25085 stained PC12 cells. The cells were 100% methanol fixed (5 min) 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 (ab25085, 1ug/ml) overnight at +4øC. The secondary antibody (green)ÿwas Alexa Fluor© 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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 1.43uM.

  • Anti-Calbindin antibody (ab25085) at 1 µg/ml + Brain (Rat) Tissue Lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 28 kDa
    Observed band size : 30 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 67 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutes

  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 28 kDa


    Exposure time : 30 seconds

参考文献

This product has been referenced in:
  • Li J  et al. Effects of age on intestinal phosphate transport and biochemical values of broiler chickens. Asian-Australas J Anim Sci 30:221-228 (2017). Chicken . Read more (PubMed: 27703131) »
  • Furukawa S  et al. Databases for technical aspects of immunohistochemistry. J Toxicol Pathol 30:79-107 (2017). IHC ; Rat . Read more (PubMed: 28190929) »

See all 12 Publications for this product

レビューと Q&A

Application
Western blot
Sample
Chicken Tissue lysate - whole (Duodenum)
Gel Running Conditions
Reduced Denaturing (gradient 4-20%)
Loading amount
10 µg
Specification
Duodenum
Blocking step
Odyssey® Blocking Buffer (PBS) as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Mr. Aurelien Brionne

Verified customer

投稿 Jul 28 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
None
Sample
Human Tissue sections (tooth)
Specification
tooth
Permeabilization
No
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

投稿 Jul 17 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Kidney)
Specification
Kidney
Fixative
Acetone
Permeabilization
No
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 21°C
Username

Abcam user community

Verified customer

投稿 Mar 30 2012

I apologize once again for my mistake in shipping you the wrong antibody. The new antibody ab49899 will be shipped on Monday and will arrive on Tuesday. Your order number is ******

I am sorry that you were having issues with ab25085 in immuno staining. I have sent you the replacement antibody ab49889, which is only listed as recognizing calbindin and also is raised against a different sequence compare to ab2085. Your new order n...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Tissue lysate - whole (Brain)
Loading amount
20 µg
Specification
Brain
Gel Running Conditions
Reduced Denaturing (4-12%)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Jul 13 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Brain)
Specification
Brain
Fixative
Paraformaldehyde
Permeabilization
Yes - Triton X-100
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Jun 01 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Kidney)
Specification
Kidney
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
Permeabilization
No
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 1%
Username

Abcam user community

Verified customer

投稿 Jul 31 2007

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

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