Overview

  • Product name

  • Description

    Rabbit polyclonal to Calbindin
  • Host species

    Rabbit
  • Specificity

    We recommend the use of RabMAb ab108404 for testing in IHC and ICC/IF for this target.
  • Tested applications

    Suitable for: IHC-Fr, IP, IHC-FoFr, WBmore 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.

    Read Abcam's proprietary immunogen policy (Peptide available as ab60452.)

  • Positive control

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

Properties

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-FoFr 1/300.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 29 kDa (predicted molecular weight: 30 kDa).

Target

Images

  • 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

  • All lanes : ab49899 at 1 µg/ml

    Lane 1 : Brain (human) tissue lysate
    Lane 2 : Brain (Mouse) Tissue lysate
    Lane 3 : Brain (Rat) Tissue lysate

    Lysates/proteins at 10 µg per lane.

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

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 30 kDa
    Observed band size: 30 kDa
    Additional bands at: 37 kDa, 51 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 30 seconds
  • All lanes : Anti-Calbindin antibody (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
    All lanes : 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.

  • 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

This product has been referenced in:

  • Schludi MH  et al. Spinal poly-GA inclusions in a C9orf72 mouse model trigger motor deficits and inflammation without neuron loss. Acta Neuropathol 134:241-254 (2017). Read more (PubMed: 28409281) »
  • Schmouth JF  et al. Non-coding-regulatory regions of human brain genes delineated by bacterial artificial chromosome knock-in mice. BMC Biol 11:106 (2013). IHC . Read more (PubMed: 24124870) »
See all 2 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Western blot
Sample
Dog Cell lysate - whole cell (hOCT2/hMATE1 double transfected MDCK cells)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Loading amount
30 µg
Specification
hOCT2/hMATE1 double transfected MDCK cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted May 10 2017

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)

Abcam user community

Verified customer

Submitted Sep 29 2015

Question
Answer




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

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Chicken Cell (chick retinal sections)
Specification
chick retinal sections
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.25% Triton

Dr. Qing Shi

Verified customer

Submitted Aug 05 2011

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

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted May 20 2011

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

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted May 19 2011

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain sections)
Specification
Brain sections
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
No

Dr. Sophie Pezet

Verified customer

Submitted May 11 2010

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