Human PLGF ELISA Kit (HUFI00021)- High Sensitivity

Key Features

	Save Time	Pre-coated 96 well plate
	Quick Start	Kit includes all necessary reagents
	Publication Ready	Reproducible and reliable results

Overview


Product Name:	Human PLGF ELISA Kit
Product Code:	HUFI00021
Size:	96 Assays
Alias:	PLGF, PGF, Placental Growth Factor, Pgf, PGF, PGFL, placenta growth factor, placental growth factor, placental growth factor, vascular endothelial growth factor-related protein, PlGF, PlGF-2, PLGFplacental growth factor-like
Detection Method:	Sandwich ELISA, Double Antibody
Application:	This immunoassay kit allows for the in vitro quantitative determination of Human Phospho Tau concentrations in serum plasma and other biological fluids.
Sensitivity:	9.375pg/ml
Range:	15.625-1000pg/ml
Storage:	4°C for 6 months
Note:	For Research Use Only

Additional Information


Recovery	Matrices listed below were spiked with certain level of Human PLGF and the recovery rates were calculated by comparing the measured value to the expected amount of Human PLGF in samples.
Linearity	The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human PLGF and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
CV(%)	Intra-Assay: CV<8% Inter-Assay: CV<10%

Kit Components

Component	Quantity	Storage
ELISA Microplate (Dismountable)	8x12 strips	4°C for 6 months
Lyophilized Standard	2	4°C/ -20°C
Sample/Standard Dilution Buffer	20ml	4°C
Biotin-labeled Antibody (Concentrated)	120ul	4°C (Protection from light)
Antibody Dilution Buffer	10ml	4°C
HRP-Streptavidin Conjugate (SABC)	120ul	4°C (Protect from light)
SABC Dilution Buffer	10ml	4°C
TMB Substrate	10ml	4°C (Protection from light)
Stop Solution	10ml	4°C
Wash Buffer (25X)	30ml	4°C
Plate Sealer	5	-

Other materials 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

Protein Information

Heading 1	Heading 2
UniProt	P49763
NCBI GenInfo Identifier	17380553
NCBI Gene ID	5228
Molecular Weight	19,338 Da
Protein Family	Placenta growth factor

Protocol

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

Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

Step	Procedure
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.

Sample Type

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.

PLGF Background

Placental growth factor (PLGF) is a protein that plays a crucial role in angiogenesis, the formation of new blood vessels. It is primarily expressed in the placenta, but it can also be found in various other tissues and cell types. Placental growth factor (PLGF) is a protein that has garnered significant attention in the field of biomedical research due to its diverse roles in angiogenesis and vascular development. Originally discovered for its expression in the placenta, PLGF has since been found to be present in various tissues and cells throughout the body. It belongs to the vascular endothelial growth factor (VEGF) family and acts as a key modulator of blood vessel formation and remodeling. PLGF interacts with specific receptors on endothelial cells, stimulating their proliferation, migration, and survival. This makes PLGF an essential factor in processes like embryonic development, tissue repair, and wound healing. Additionally, PLGF has been implicated in pathological conditions such as cancer, where it can contribute to tumor angiogenesis and metastasis. PLGF functions in angiogenesis, respiratory development, and spermatogenesis. PLGF is used as a marker for human embryonic stem cells and is found at higher levels in seminal plasma than platelets or blood

PLGF Structure

The human PlGF gene is situated on chromosome 14q14 and encodes four distinct isoforms of PlGF. This protein is released as a glycosylated homodimer. Among the isoforms, PlGF-1 and PlGF-3 are diffusible, while PlGF-2 and PlGF-4 contain heparin binding domains. PlGF-1 and PlGF-2 are the most prevalent forms, and during pregnancy, their secretion shows a strong correlation, indicating a shared regulatory mechanism. The presence of a heparin binding domain suggests that PlGF-2 and PlGF-4 remain associated with cell membranes, exerting their effects in an autocrine manner. Conversely, the diffusible forms of PlGF likely impact target cells in a paracrine fashion.

PLGF ELISA Kit FAQs

What is the PLGF ELISA Kit used for?

PLGF is a key regulator of angiogenesis, the formation of new blood vessels. ELISA kits can be used to quantify PLGF levels in different tissues or cell cultures to study its involvement in angiogenesis and vascular development. It can also be used in measurement of PLGF levels in maternal serum or amniotic fluid, aiding in the assessment of placental function, monitoring of pregnancy complications, and prediction of preeclampsia and other gestational disorders.

What are the advantages of using the PLGF ELISA Kit?

The PLGF ELISA Kit offers several advantages, including high sensitivity, accuracy, and reproducibility. It provides a user-friendly and reliable method to quantify PLGF levels in biological specimens, allowing for precise measurements and robust data analysis.

What sample types are compatible with PLGF ELISA Kit?

The PLGF ELISA Kit is compatible with various sample types, including serum, plasma, cell lysates, and tissue homogenates. It provides flexibility in sample selection, allowing researchers to analyze PLGF levels in different biological matrices.

What are the storage requirements with PLGF ELISA Kit?

The PLGF ELISA Kit components should be stored according to the instructions provided in the kit manual. Generally, it is recommended to store the kit components at the recommended temperature to ensure their stability and optimal performance.

What should I do if my assay results are not optimal?

If you encounter any issues or have suboptimal assay results, we recommend contacting our dedicated support team for assistance. They will be available to provide troubleshooting guidance, answer your questions, and ensure you achieve the best possible results with the PLGF ELISA Kit.

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Human PLGF ELISA Kit

Description