Anti-Calbindin antibody - Neuronal Marker (ab49899)

Overview

  • Product nameAnti-Calbindin antibody - Neuronal Marker
    See all Calbindin primary antibodies
  • Description
    Rabbit polyclonal to Calbindin - Neuronal Marker
  • Tested applicationsSuitable for: IHC-Fr, IP, IHC-P, IHC-FoFr, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Rabbit, Hamster, Cow, Pig
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Mouse Calbindin.

    (Peptide available as ab60452.)

  • Positive control
    • This antibody gave a positive signal in the following tissues: Kidney (Mouse), Kidney (Human), Brain (Rat)

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab49899 in the following tested applications.

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

Application Abreviews Notes
IHC-Fr 1/200.
IP Use at an assay dependent concentration.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-FoFr 1/300.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 29 kDa (predicted molecular weight: 30 kDa).
ICC/IF Use a concentration of 5 µg/ml.

Target

Anti-Calbindin antibody - Neuronal Marker images

  • All lanes : Anti-Calbindin antibody - Neuronal Marker (ab49899) at 1 µg/ml

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

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 30 kDa
    Observed band size : 30 kDa
    Additional bands at : 150 kDa. We are unsure as to the identity of these extra bands.
  • Immunohistochemistical detection of Calbindin using antibody ab49899 on PFA perfusion-fixed frozen rat brain sections (30um). Antigen retrieval step: None Primary Antibody diluted at 1/300 and incubated for 18 hours @ 20°C in PBS + 0.3 % Triton X100. Secondary Antibody: Goat anti-rabbit Alexa Fluor® 488 (1/1000). ab49899 was used on sections of rat brain in free floating IHC; expected staining was observed on rat brain. The submitted images show staining obtained in the cerebral cortex (A,B). The images were taken using the objective X5 (A) and X40 (B) and show the neuronal profiles (arrows) and neuronal processes.

    See Abreview

  • ICC/IF image of ab49899 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 (ab49899, 5µg/ml) overnight at +4°C. The secondary antibody (green) was 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 1.43µM.
  • IHC image of Calbindin staining in rat brain 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 ab49899, 1µg/ml, 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.
  • Calbindin - Neuronal Marker was immunoprecipitated using 0.5mg Mouse kidney whole 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, Mouse kidney whole 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab49899.
    Secondary: Clean blot (HRP conjugate) at 1/1000 dilution.
    Band: 30kDa: Calbindin - Neuronal Marker.

References for Anti-Calbindin antibody - Neuronal Marker (ab49899)

ab49899 has not yet been referenced specifically in any publications.

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Guinea pig Tissue sections (Brain)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer
Permeabilization No
Specification Brain
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: RT°C
Fixative neutral buffered formalin (NBF)
Username

Abcam user community

Verified customer

Submitted Sep 29 2015




The image is showing the cerebral cortex of the rat brain.

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Chicken Cell (chick retinal sections)
Specification chick retinal sections
Fixative Paraformaldehyde
Permeabilization Yes - 0.25% Triton
Username

Dr. Qing Shi

Verified customer

Submitted Aug 05 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (Brain section)
Specification Brain section
Fixative Paraformaldehyde
Permeabilization Yes - 0.3% TritonX in 0.1% PBS
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted May 20 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (Primary Hippocampal culture)
Specification Primary Hippocampal culture
Fixative Methanol
Permeabilization Yes - 0.3% TritonX in 0.1% PBS
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted May 19 2011

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 sections)
Specification Brain sections
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization No
Username

Dr. Sophie Pezet

Verified customer

Submitted May 11 2010

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