Calretinin antibody (Calbindin 2) (Center)

Details for Product anti-CALB2 Antibody No. ABIN2855596
Antigen
  • CAB29
  • CAL2
  • CR
  • calb2l
  • wu:fq18e08
  • zgc:73115
  • calb2
  • wu:fq17g09
  • zgc:73099
  • calbindin 2
  • calbindin 2a
  • calbindin 2b
  • CALB2
  • Calb2
  • calb2a
  • calb2b
Epitope
Center
15
15
9
8
5
5
4
4
3
3
2
2
2
2
1
1
1
1
1
Reactivity
Dog (Canine), Human, Mouse (Murine)
168
99
95
11
6
5
4
4
4
3
2
2
2
2
1
Host
Rabbit
137
37
12
4
1
Clonality
Polyclonal
Conjugate
This Calretinin antibody is un-conjugated
8
5
5
5
5
5
5
3
2
1
1
1
1
1
1
1
Application
Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Western Blotting (WB)
110
74
47
47
21
13
13
10
10
8
7
6
6
2
1
1
1
1
1
1
Options
'Independent Validation' Badge
Antigen Calretinin
Lot Number 39876
Method validated Immunohistochemistry
Positive Control

Adult (24 months) mouse and dog brain

Negative Control

We incubated slices overnight with the blocking solution only and then processed them with the secondary antibody.

Notes

ABIN2855596 works in IHC-P with microwave antigen retrieval pretreatment at an optimal concentration of 1:200.

Primary Antibody ABIN2855596
Secondary Antibody goat anti-rabbit AF488-conjugated antibody (Invitrogen by Thermo Fisher Scientific, A11034, lot 1971418)
Protocol
  • Obtain dog samples after autopsy and immersion-fix in buffered formaldehyde for 16h.
  • Perfuse mice with paraformaldehyde 4% in 0.1M phosphate buffer pH 7.4 and post-fix in the same fixative for an additional 2h at RT.
  • Wash, dehydrate, and embed samples in paraffin wax.
  • Cut with a microtome into 6µm sections and mount on glass slides.
  • After paraffin removal, process sections by microwave antigen retrieval for 10min (95-100°C) in 10mM sodium citrate buffer (pH 6.0).
  • Let sections cool for 10min.
  • Wash sections under running water for 5-10min.
  • Wash sections 2x 5min with distilled water.
  • Wash sections 5min with 0.01M PBS.
  • Incubate sections in PBS containing 1% albumin from chicken egg white (Sigma, A5378) for 1h at RT to block non-specific binding sites.
  • Incubate sections with primary rabbit anti-Calretinin antibody (antibodies-online, ABIN2855596, lot 39876) diluted 1:200 in PBS-BSA-PLL at 4°C.
  • Wash sections 3x 5min with 0.01M PBS.
  • Incubate sections with secondary goat anti-rabbit AF488-conjugated antibody (Invitrogen by Thermo Fisher Scientific, A11034, lot 1971418) diluted 1:500 in 0.1M PBS for 1h at RT.
  • Wash sections 3x 5min with 0.01M PBS.
  • Mount specimens in Fluoroshield (Sigma, F6182, lot MKCB0153V).
  • Acquire images with a fluorescence microscope and appropriate filter settings for AF488, e.g. Leica DM 6000B fluorescence microscope equipped with the manufacturer’s FITC filter set and digital camera at magnifications of 20x, 40x, and 63x. Parameters for image acquisition (Exposure 2.2s, Gain 2.3x, Saturation 1.5, Gamma 1.10) unchanged for all images.
Experimental Notes

For the incubation with the primary antibody ABIN2669973 1:100, 1:500, and 1:1000 dilutions were also tested.

Validation Images
Immunohistochemistry validation image for anti-Calbindin 2 (CALB2) (Center) antibody (ABIN2855596) Staining of dog and mouse brain sections using ABIN2855596. A-C: Specific staining of...
Immunogen Recombinant protein encompassing a sequence within the center region of human Calretinin. The exact sequence is proprietary.
Isotype IgG
Cross-Reactivity Chicken, Pig (Porcine), Rat (Rattus), Cow (Bovine)
Cross-Reactivity (Details) Chicken (89 %), Pig (96 %), Rat (98 %), Bovine (97 %)
Characteristics Rabbit Polyclonal antibody to Calretinin (calbindin 2)
Calretinin antibody
Purification Purified by antigen-affinity chromatography.
Plasmids, Primers & others Plasmids, Primers & others Calretinin products on genomics-online (e.g. as negative or positive controls)
Antigen
Alternative Name Calretinin (CALB2 Antibody Abstract)
Background This gene encodes an intracellular calcium-binding protein belonging to the troponin C superfamily. Members of this protein family have six EF-hand domains which bind calcium. Three alternatively spliced transcript variants that encode different proteins have been described.

Molecular Weight 32 kDa
Gene ID 794
Application Notes Suggested dilution Reference ICC/IF 1:100-1:1000* IHC (Formalin-fixed paraffin-embedded sections) 1:100-1:1000* Western blot 1:500-1:3000* Not tested in other applications. *Optimal dilutions/concentrations should be determined by the researcher.Suggested dilutionReferenceICC/IF1:100-1:1000* IHC (Formalin-fixed paraffin-embedded sections)1:100-1:1000* Western blot1:500-1:3000*
Comment

Positive Control: A431 , mouse brain , human CALB2-transfected 293T cells

Restrictions For Research Use only
Format Liquid
Concentration 0.75 mg/mL
Buffer 0.1M Tris, 0.1M Glycine, 10 % Glycerol ( pH 7). 0.01 % Thimerosal was added as a preservative.
Preservative Thimerosal (Merthiolate)
Precaution of Use This product contains Thimerosal (Merthiolate): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage -20 °C
Storage Comment Keep as concentrated solution. Aliquot and store at -20°C or below. Avoid multiple freeze-thaw cycles.
Supplier Images
Immunofluorescence (IF) image for anti-Calbindin 2 (CALB2) (Center) antibody (ABIN2855596) ICC/IF Image Immunofluorescence analysis of paraformaldehyde-fixed A431, using Calret...
Western Blotting (WB) image for anti-Calbindin 2 (CALB2) (Center) antibody (ABIN2855596) WB Image Sample (50 ug of whole cell lysate) A: mouse brain 12% SDS PAGE antibody dil...
Immunohistochemistry (IHC) image for anti-Calbindin 2 (CALB2) (Center) antibody (ABIN2855596) IHC-P Image Calretinin antibody detects Calretinin protein at cytosol on mouse fore b...
Western Blotting (WB) image for anti-Calbindin 2 (CALB2) (Center) antibody (ABIN2855596) WB Image Calretinin antibody detects CALB2 protein by Western blot analysis. A. 30 ug...
Immunohistochemistry (IHC) image for anti-Calbindin 2 (CALB2) (Center) antibody (ABIN2855596) IHC-P Image Immunohistochemical analysis of paraffin-embedded Mouse retina, using Cal...
Western Blotting (WB) image for anti-Calbindin 2 (CALB2) (Center) antibody (ABIN2855596) WB Image Sample (30 ug of whole cell lysate) A: A431 , 12% SDS PAGE Calretinin antibo...
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