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Mouse Calbindin / CALB1 ELISA Kit

SKU:
MOFI00345
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P12658
Sensitivity:
18.75pg/ml
Range:
31.25-2000pg/ml
ELISA Type:
Sandwich
Synonyms:
Calb1, CAB27, Vitamin D-dependent calcium-binding protein, avian-type, Calbindin D28, D-28K
Reactivity:
Mouse
$779
Frequently bought together:

Description

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Mouse Calbindin/CALB1 ELISA Kit

The Mouse Calbindin (CALB1) ELISA Kit is specifically designed for the precise quantification of calbindin levels in mouse serum, plasma, and tissue lysates. With a high level of sensitivity and specificity, this kit ensures accurate and consistent results, making it ideal for various research applications.Calbindin, encoded by the CALB1 gene, is a calcium-binding protein that plays a crucial role in the regulation of calcium homeostasis and neuronal function. It is involved in various physiological processes, including muscle contraction, neurotransmitter release, and neuronal survival.

Abnormalities in calbindin expression have been linked to neurological disorders, such as Alzheimer's disease, Parkinson's disease, and epilepsy. By studying calbindin levels using this ELISA kit, researchers can gain valuable insights into the pathophysiology of these conditions and potentially identify new therapeutic targets.Overall, the Mouse Calbindin (CALB1) ELISA Kit is a valuable tool for investigating the role of calbindin in health and disease, providing researchers with reliable data to advance their scientific studies.

Product Name:Mouse Calbindin / CALB1 ELISA Kit
Product Code:MOFI00345
Size:96 Assays
Alias:Calb1, CAB27, Vitamin D-dependent calcium-binding protein, avian-type, Calbindin D28, D-28K
Detection Method:Sandwich ELISA
Application:This immunoassay kit allows for the in vitro quantitative determination of Mouse Calb1 concentrations in serum plasma and other biological fluids.
Sensitivity:18.75pg/ml
Range:31.25-2000pg/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery: Matrices listed below were spiked with certain level of Mouse Calb1 and the recovery rates were calculated by comparing the measured value to the expected amount of Mouse Calb1 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)88-10594
EDTA plasma(n=5)86-10396
UFH plasma(n=5)85-9590
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Mouse Calb1 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)85-102%89-98%86-102%
EDTA plasma(n=5)83-96%82-97%87-94%
UFH plasma(n=5)81-96%80-95%83-96%
Intra Assay:CV <8%
Inter Assay:CV <10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8-12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir
UniprotP12658
UniProt Protein Function:CALB1: Buffers cytosolic calcium. May stimulate a membrane Ca(2+)-ATPase and a 3',5'-cyclic nucleotide phosphodiesterase. Interacts with RANBP9. Belongs to the calbindin family.
UniProt Protein Details:

Protein type:Calcium-binding

Chromosomal Location of Human Ortholog: 4 A2|4 6.66 cM

Cellular Component: axon; cell soma; cytoplasm; cytosol; dendrite; intracellular; neuron projection; nucleus; synapse; terminal button

Molecular Function:calcium ion binding; zinc ion binding

Biological Process: cytosolic calcium ion homeostasis; locomotory behavior; long-term memory; regulation of synaptic plasticity; retina development in camera-type eye; short-term memory

UniProt Code:P12658
NCBI GenInfo Identifier:6753242
NCBI Gene ID:12307
NCBI Accession:NP_033918.1
UniProt Secondary Accession:P12658,Q545N6,
UniProt Related Accession:P12658
Molecular Weight:29,994 Da
NCBI Full Name:calbindin
NCBI Synonym Full Names:calbindin 1
NCBI Official Symbol:Calb1  
NCBI Official Synonym Symbols:CB; Calb; Calb-1; Brain-2  
NCBI Protein Information:calbindin
UniProt Protein Name:Calbindin
UniProt Synonym Protein Names:Calbindin D28; D-28K; PCD-29; Spot 35 protein; Vitamin D-dependent calcium-binding protein, avian-type
Protein Family:Coniferyl aldehyde dehydrogenase
UniProt Gene Name:Calb1  

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

StepProcedure
1. Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3. Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4. Add 0.1 ml of properly diluted sample (Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
6. Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7. Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9. Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10. Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11. Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12. Add 90 µL of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13. Add 50 µL of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type Protocol
Serum:

If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

Plasma:

Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 - g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.

Urine & Cerebrospinal Fluid:

Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.

Cell culture supernatant:

Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.

Cell lysates:

Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20°C.

Tissue homogenates:

The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.

Tissue lysates:

Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.

Breast Milk:

Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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