High Sensitivity Human Growth Hormone (H-hGH) ELISA Kit

Name: High Sensitivity Human Growth Hormone (H-hGH) ELISA Kit
SKU: DEIA449
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

COA

SDS

Datasheet

Summary

Sample

Sreum

Species Reactivity

Human

Detection Method

sELISA

Intended Use

This high-sensitivity Human Growth Hormone ELISA Kit is to be used for the in vitro quantitative determination of human growth hormone (hGH) concentrations in serum. This kit is intended FOR LABORATORY RESEARCH USE ONLY, and is not to be used in diagnostic or therapeutic procedures.

Contents of Kit

All reagents provided are stored at 2-8°C. Refer to expiration date on the label.
1. MICROTITER PLATE (96 wells): Pre-coated with anti-human hGH monoclonal antibody.
2. CONJUGATE (12 mL): Anti-hGH monoclonal antibody conjugated to horseradish peroxidase (HRP) with preservative. Ready to use.
3. hGH STANDARD - 12 ng/mL (1 vial): Lypholized human hGH in a buffered protein base with preservative that will contain 12 ng/mL after reconstitution.
4. hGH STANDARD - 6 ng/mL (1 vial): Lypholized human hGH in a buffered protein base with preservative that will contain 6 ng/mL after reconstitution.
5. hGH STANDARD - 2 ng/mL (1 vial): Lypholized human hGH in a buffered protein base with preservative that will contain 2 ng/mL after reconstitution.
6. hGH STANDARD - 0.8 ng/mL (1 vial): Lypholized human hGH in a buffered protein base with preservative that will contain 0.8 ng/mL after reconstitution.
7. hGH STANDARD - 0.2 ng/mL (1 vial): Lypholized human hGH in a buffered protein base with preservative that will contain 0.2 ng/mL after reconstitution.
8. hGH STANDARD - 0 ng/mL (1 vial): Lypholized buffered protein base with preservative that will contain 0ng/mL after reconstitution.
9. SAMPLE DILUENT (7 mL): Animal serum in buffer with 0.05% proclin-300.
10. SUBSTRATE A (10 mL): Buffered solution with H₂O₂.
11. SUBSTRATE B (10 mL): Buffered solution with TMB.
12. STOP SOLUTION (14 mL): 2N H₂SO₄. Caution: Caustic Material!
13. WASH BUFFER (20 ×) (60 mL): 20-fold concentrated solution of buffered surfactant.

Storage

All reagents provided are stored at 2-8°C.

Performance Characteristics

CALIBRATION:
This immunoassay is calibrated against NIBSC/WHO, 98/574, hGH.
HOOK EFFECT
In this assay, no hook effect is observed.
EXPECTED NORMAL VALUES
Each laboratory must establish its own normal range based on patient population. The results provided below are based on random selected outpatient clinical laboratory samples:

Sensitivity

The minimal detectable concentration of hGH by this assay is estimated to be 0.1 ng/mL.

General Description

Human growth hormone (GH) is a 22kDa monomeric protein produced and stored in somatotrophs in the anterior pituitary gland. GH is released from the pituitary into the bloodstream in a pulsatile manner under the regulatory control of hypothalamic somatostatin (SS) and GH-releasing factor (GHRF) [1]. The timing and frequency of GH release appears to be regulated by somatostatin, while the amplitude of GH release is determined by GHRF. A minor fraction (~10%) of GH in circulation exists in a smaller 20 kDa form [2]. GH has profound effects on tissue growth and metabolism, which is thought to be mediated through GH-dependent production of IGF-I and IGF-II, and their associated binding proteins. GH apparently stimulates IGF production after binding to specific cell surface receptors in the liver and, possibly, other tissues. Almost 50% of GH in circulation is bound to a high affinity GH binding protein (GHBP), which represents the extracellular domain of the GH receptor. Deficient GH secretion can occur in a number of clinical conditions [3]. However, evaluation of GH deficiency is complicated by the episodic nature of GH secretion and low circulating levels. A variety of physiologic and pharmacologic stimuli have been used to stimulate pituitary GH release during testing and failure to achieve a normal serum GH level in response to at least 2 GH stimulation or provocative tests is considered to be a diagnostic of GH deficiency [4]. The definition of a normal serum GH response is controversial, although published values generally range from 5 to 10 ng/mL. GH excess (or acromegaly) can be caused either by direct GH hypersecretion or GH excess secondary to GHRF hypersecretion.

Standard Curve

Results of a typical standard run of hGH ELISA are shown. Any variation in operator, pipetting and washing technique, incubation time or temperature, and kit age can cause variation in result. The following examples are for the purpose of illustration only, and should not be used to calculate unknowns. Each users should obtain their own standard curve.
EXAMPLE: Results of a typical standard run are shown below:

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Documents

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QA & Reviews

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References

Tannase Sequence from a Xerophilic Aspergillus niger Strain and Production of the Enzyme in Pichia pastoris

MOLECULAR BIOTECHNOLOGY

Authors: Antonio Fuentes-Garibay, Jose; Noe Aguilar, Cristobal; Rodriguez-Herrera, Raul; Guerrero-Olazaran, Martha; Maria Viader-Salvado, Jose

Abstract

Tannin acyl hydrolases, or tannases (EC 3.1.1.20), are enzymes with potential biotechnological applications. In this work, we describe the gene and amino acid sequences of the tannase from Aspergillus niger GH1. In addition, we engineered Pichia pastoris strains to produce and secrete the enzyme, and the produced tannase was characterized biochemically. The nucleotide sequence of mature tannase had a length of 1,686 bp, and encodes a protein of 562 amino acids. A molecular model of mature A. niger GH1 tannase showed the presence of two structural domains, one with an alpha/beta-hydrolase fold and one lid domain that covers the catalytic site, likely being residues Ser-196, Asp-448, and His-494 the putative catalytic triad, which are connected by a disulfide bond between the neighboring cysteines, Cys-195 and Cys-495. A 120-ml shake flask culture with a constructed recombinant P. pastoris strain showed extracellular tannase activity at 48 h induction of 0.57 U/ml. The produced tannase was N-glycosylated, consisted of two subunits, likely linked by a disulfide bond, and had an optimum pH of 5.0 and optimum temperature of 20 degrees C. These biochemical properties differed from those of native A. niger GH1 tannase. The recombinant tannase could be suitable for food and beverage applications.

Metatranscriptomic Analyses of Plant Cell Wall Polysaccharide Degradation by Microorganisms in the Cow Rumen

APPLIED AND ENVIRONMENTAL MICROBIOLOGY

Authors: Dai, Xin; Tian, Yan; Li, Jinting; Su, Xiaoyun; Wang, Xuewei; Zhao, Shengguo; Liu, Li; Luo, Yingfeng; Liu, Di; Zheng, Huajun; Wang, Jiaqi; Dong, Zhiyang; Hu, Songnian; Huang, Li

Abstract

The bovine rumen represents a highly specialized bioreactor where plant cell wall polysaccharides (PCWPs) are efficiently deconstructed via numerous enzymes produced by resident microorganisms. Although a large number of fibrolytic genes from rumen microorganisms have been identified, it remains unclear how they are expressed in a coordinated manner to efficiently degrade PCWPs. In this study, we performed a metatranscriptomic analysis of the rumen microbiomes of adult Holstein cows fed a fiber diet and obtained a total of 1,107,083 high-quality non-rRNA reads with an average length of 483 nucleotides. Transcripts encoding glycoside hydrolases (GHs) and carbohydrate binding modules (CBMs) accounted for similar to 1% and similar to 0.1% of the total non-rRNAs, respectively. The majority (similar to 98%) of the putative cellulases belonged to four GH families (i.e., GH5, GH9, GH45, and GH48) and were primarily synthesized by Ruminococcus and Fibrobacter. Notably, transcripts for GH48 cellobiohydrolases were relatively abundant compared to the abundance of transcripts for other cellulases. Two-thirds of the putative hemicellulases were of the GH10, GH11, and GH26 types and were produced by members of the genera Ruminococcus, Prevotella, and Fibrobacter. Most (similar to 82%) predicted oligosaccharide-degrading enzymes were GH1, GH2, GH3, and GH43 proteins and were from a diverse group of microorganisms. Transcripts for CBM10 and dockerin, key components of the cellulosome, were also relatively abundant. Our results provide metatranscriptomic evidence in support of the notion that members of the genera Ruminococcus, Fibrobacter, and Prevotella are predominant PCWP degraders and point to the significant contribution of GH48 cellobiohydrolases and cellulosome-like structures to efficient PCWP degradation in the cow rumen.