Human Cathepsin H ELISA Kit PicoKine®

Cathepsin H ELISA kit for Human

Human Cathepsin H ELISA Kit PicoKine™ (96 Tests). Quantitate Human CTSH in cell culture supernatants, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml .

Product Info Summary

SKU: EK0674
Size: 96 wells/kit, with removable strips.
Reactive Species: Human
Application: ELISA
Sample Types: cell culture supernatants, serum and plasma (heparin, EDTA).

Product Name

Human Cathepsin H ELISA Kit PicoKine®

View all Cathepsin H ELISA kits

SKU/Catalog Number

EK0674

Size

96 wells/kit, with removable strips.

*Question: How many samples can I assay/run in this kit?

Description

Human Cathepsin H ELISA Kit PicoKine™ (96 Tests). Quantitate Human CTSH in cell culture supernatants, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml .

Storage & Handling

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Cite This Product

Human Cathepsin H ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0674)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: CHO; Immunogen sequence: A23-V335

Sensitivity

<10 pg/ml

Assay Range

156 pg/ml - 10,000 pg/ml

Standard Dilution Instructions

serial dilution instructions image

Add 100ul of sample diluent in well #2-#8. Add 200ul standard stock solution to well #1, and serial dilute for well #2-#7 to make a standard curve row. Leave well #8 as blank See datasheet of EK0674 for more details

Cross-reactivity

There is no detectable cross-reactivity with other relevant proteins.

Reactive Species

EK0674 is reactive to CTSH in Human samples

Validated Sample Types

cell culture supernatants, serum and plasma (heparin, EDTA).

Application Guarantee

EK0674 is guaranteed for ELISA in Human by Boster Guarantee

See how Boster Bio validate our ELISA kits: ELISA Validation Information

Background of Cathepsin H

Cathepsin H is a protein that in humans is encoded by the CTSH gene. The protein encoded by this gene gene is a lysosomal cysteine proteinase important in the overall degradation of lysosomal proteins. It is composed of a dimer of disulfide-linked heavy and light chains, both produced from a single protein precursor. The encoded protein, which belongs to the peptidase C1 protein family, can act both as an aminopeptidase and as an endopeptidase. Increased expression of this gene has been correlated with malignant progression of prostate tumors. Alternate splicing of this gene results in multiple transcript variants encoding different isoforms.

Kit Components

Catalog Number Description Quantity
EK0674-CAP Anti-Human CTSH Pre-coated 96-well strip microplate 1
EK0674-ST Human CTSH Standard 2 vials, 10 ng/tube
EK0674-DA Human CTSH Biotinylated antibody (100x) 100ul
AR1103 Avidin-Biotin-Peroxidase Complex (100x) 100ul
AR1106-1 Sample Diluent 30ml
AR1106-2 Antibody Diluent 12ml
AR1106-3 Avidin-Biotin-Peroxidase Diluent 12ml
AR1104 Color Developing Reagent (TMB) 10ml
AR1105 Stop Solution 10ml
AR1106-5 Wash Buffer (25x) 20ml
PLA-SEA Adhesive plate sealers 4

*The kit components are not available for individual purchase.

Materials Required But Not Included In Kit

  • Microplate Reader capable of reading absorbance at 450nm.
  • Incubator.
  • Automated plate washer (optional).
  • Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
  • Multichannel pipettes are recommended for large amount of samples.
  • Deionized or distilled water.
  • 500ml graduated cylinders.
  • Test tubes for dilution.

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Validation Standard Curve O.D. At 450nm

Concentration (pg/ml)015631262512500025000500010000
O.D.0.1400.2110.2830.3730.6111.0251.6222.382

Data Example Images

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Human Cathepsin H ELISA Kit PicoKine® within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.

Intra-Assay PrecisionInter-Assay Precision
Sample123123
n161616242424
Mean (pg/ml)3729874039539510223714
Standard deviation16.7468.11622.91922.9180.73185.7
CV (%)4.5%6.9%4.1%5.8%7.9%5%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of Cathepsin H in ELISA kits from four different production batches/lots.

LotsLot 1 (pg/ml)Lot 2 (pg/ml)Lot 3 (pg/ml)Lot 4 (pg/ml)Mean (pg/ml)Standard DeviationCV (%)
Sample 137231531833533522.686.7%
Sample 298798710841153105270.126.6%
Sample 340954519462242214364214.124.9%
*number of samples for each test n=16.

Gene/Protein Information For CTSH (Source: Uniprot.Org, NCBI)

Gene Name

CTSH

Full Name

Pro-cathepsin H

Weight

37.394kDa

Superfamily

peptidase C1 family

Alternative Names

ACC-4; ACC-5; aleurain; cathepsin B3; cathepsin BA; Cathepsin H; CPSB; CTSH; DKFZp686B24257; EC 3.4.22; EC 3.4.22.16; MGC1519; minichain; N-benzoylarginine-beta-naphthylamide hydrolase CTSH ACC-4, ACC-5, ACC4, ACC5, CPSB cathepsin H pro-cathepsin H|N-benzoylarginine-beta-naphthylamide hydrolase|aleurain|cathepsin B3|cathepsin BA

*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".

For more info on CTSH, check out the CTSH Infographic

CTSH infographic

We have 30,000+ of these available, one for each gene! check them out.

In this infographic you will see the following information for CTSH: database IDs, super-family, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact us [email protected].

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10 Customer Q&As for Human Cathepsin H ELISA Kit PicoKine®

Question

Q: how to thaw whole blood sample for Cathepsin H ELISA after freezing?

T. Hill

Verified customer

Asked: 2020-06-17

Answer

A: do not freeze and thaw whole blood. erythrocytes are fragile and, if frozen and thawed, will undergo hemolysis rendering the samples useless. To keep your blood samples to test Cathepsin H for a later time, you should let the blood clot in glass tubes and collect the serum to freeze for later use.

Boster Scientific Support

Answered: 2020-06-17

Question

Q: can you suggest the dilution ratio of serum samples for detection of Cathepsin H in Human serum? I am trying to measure a multiple analytes and it requires 100ul of diluted samples for each well. We have low serum volumes so we like to dilute as much as possible.

Verified Customer

Verified customer

Asked: 2019-06-14

Answer

A: having little idea about the physiological or pathological context of your samples we cannot recommend a dilution ratio without performing a pilot test with your samples. Here is how you can perform a pilot study on your own: perform a serial dilution of your samples on the Cathepsin H ELISA kit to make sure you have a linear ascending curve followed by a plateau, which signifies the samples saturating the detection limit of the kit. Then you can pick the dilution ratios from samples in the linear part of the curve as your experimental dilution ratio.
If you are interested in using our ELISA service, you can also send us your sample and we will take care of everything for you. You can check our service details here: bosterbio.com/services/assay-services/ELISA-testing-service
Since you mentioned you have limited samples, our cost effective multiplex ELISA service would fit perfectly for your needs, where we can generate dozens of data points using as little as 25ul sample volume. Information on this service is also in the above link.

Boster Scientific Support

Answered: 2019-06-14

Question

Q: what is the protocol regarding preparation of cell lysates prepared for use in Picokine® ELISA kits?

D. Chakrabarti

Verified customer

Asked: 2019-05-19

Answer

A: for those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate can be found in the product insert. Components in lysate and lysis buffer can change immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.

Boster Scientific Support

Answered: 2019-05-19

Question

Q: Are Boster Bio recombinant proteins and antibodies sterile?

Verified Customer

Verified customer

Asked: 2019-04-06

Answer

A: although the vials are bottled using aseptic techniques, heat-treated vials, and sterile stock solutions, they are not considered or guaranteed to be sterile. If sterile material is a must for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.

Boster Scientific Support

Answered: 2019-04-06

Question

Q: how many samples can be assayed in a Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2018-11-20

Answer

A: The Picokine® ELISA Kits will generally run a 7-point standard curve, non-specific binding wells, and 39 samples in duplicate. this may might differ by kit so please refer to each datasheet for details.

Boster Scientific Support

Answered: 2018-11-20

Question

Q: What is the optimal O.D. value for Cathepsin H ELISA kit? I used your Cathepsin H ELISA on serum samples. For my positive control, I received an O.D. value of 0.826, while my negative control received a value of 0.136. I obtained both of these controls from the ELISA kit, where your kit's typical data shows O.D. values much higher than my positive control and your background is lower. My samples O.D. values are around 0.225 and the highest is only 0.357. can I consider these samples contain Cathepsin H even though the O.D. values are not very high?

Verified Customer

Verified customer

Asked: 2018-09-23

Answer

A: The absolute O.D. values may change according to incubation time. The more you incubate the higher the O.D. values are going to be. an important assessment should be is whether your sample O.D. values are statistically significantly higher than your blank values. in the above example, you could extend your development time in the substrate incubation step to obtain higher O.D. values, as long as your negative controls' O.D. values are not increasing faster in relation to your positive controls. normally, a sample with O.D. value 2 standard deviations higher than your negative controls can be considered positive. We calculate the sensitivity of this ELISA kit by changeing cutoff O.D. value, calculated as the average of 20 negative controls plus 2 standard deviations of the 20 negative controls, into a concentration. in other words, when we claim this Cathepsin H ELISA kit to have sensitivity of 10pg/ml , that means the minimum amount of Cathepsin H that can be declared/interpreted as positive by the above standard is 10pg/ml .

Boster Scientific Support

Answered: 2018-09-23

Question

Q: if the enzyme conjugated Cathepsin H antibodies are mixed with the substrate, will that change the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the Cathepsin H antigen?

J. Hill

Verified customer

Asked: 2018-07-03

Answer

A: The enzyme is always active. Avoid contaminating the substrate with enzyme prior to the incubation otherwise it compromises the assay with false positive signal.

Boster Scientific Support

Answered: 2018-07-03

Question

Q: can I use citrate plasma as samples in Human Cathepsin H Picokine® ELISA Kit (Catalog # EK0674)?

Verified Customer

Verified customer

Asked: 2017-12-08

Answer

A: Chelating agents such as EDTA, Heparin and Citrate can irreversibly bind metal ions from the functional domain of Cathepsin H causing degradation of its protein structure. Cathepsin H may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes required for detection and block the antigen antibody reaction. We have tested the Cathepsin H ELISA, treating samples with different anticoagulants and decided that heparin or EDTA can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.

Boster Scientific Support

Answered: 2017-12-08

Question

Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for measurement Cathepsin H using the EK0674 Human Cathepsin H Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2015-04-10

Answer

A: Boster Bio does not evaluate sample stability. Variations in sample collection, processing, and storage may alter the stabilityof samples. It is recommend to assay sample immediately after collection when possible, or aliquot into single use volumes and store samples frozen. avoid repeat freeze-thaw cycles with the stored samples to prevent protein degradation.

Boster Scientific Support

Answered: 2015-04-10

Question

Q: is there any online tool I can use to streamline the data analysis for my ELISA results?

Verified Customer

Verified customer

Asked: 2014-02-12

Answer

A: We have a web based ELISA curve fitting (4pl) and data analysis tool. Please give it a try: bosterbio.com/biology-research-tools/ELISA-data-analysis-online. You can also consult our article on ELISA data analysis: bosterbio.com/ELISA-data-analysis-instructions

Boster Scientific Support

Answered: 2014-02-12

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