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AB326899

Human VEGF ELISA Kit, Chemiluminescent

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Human VEGF ELISA Kit, Chemiluminescent is a 55-min single-wash SimpleStep Ignite™ ELISA for the rapid quantification of Human VEGF with higher sensitivity and broader measuring range.

- Single 30 min incubation and single-wash step for results in less than an hour.
- Chemiluminescent detection allows a broad dynamic range: 5.49 - 4000 pg/ml with a sensitivity of 1.93 pg/mL.

- Patent pending

View Alternative Names

VEGF, VEGFA, L-VEGF, Vascular permeability factor, VPF

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Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)
  • sELISA

Supplier Data

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)

Interpolated concentrations of native VEGF in cell culture supernatant samples.

The concentrations of VEGF were measured in duplicates, interpolated from the VEGF standard curves and corrected for sample dilution. Undiluted samples are as follows : HepG2 5%, A431 5%, MDA-MB-435S 12.5%, PC-3 20%, A549 50%, and PBMC 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean VEGF concentration was determined to be 5027 pg/mL in neat HepG2 supernatant, 7356 pg/mL in neat A431 supernatant (4 Day), 5918 pg/mL in neat MDA-MB-435S supernatant, 1495 pg/mL in neat PC-3 (2 Day), 603.4 pg/mL in neat A549 supernatant, and 405.3 pg/mL in neat PBMC supernatant (PHA-M, 5 Day).

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)
  • sELISA

Supplier Data

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)

Other species reactivity was determined by measuring a 500 pg/mL protein load of various species recombinant VEGF proteins.

Protein concentrations were interpolated from the human standard curve, and graphing the interpolated concentrations (mean +/- SD, n=2).

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)
  • sELISA

Supplier Data

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)

Comparison of VEGF in PHA-M stimulated and unstimulated human PBMC cell culture supernatants stimulated for different durations.

Human PBMC (seeded at 10x106/mL) were cultured for 2 or 5 days in the presence or absence of 1.5% PHA-M. The concentrations of VEGF were measured in three different dilutions of the supernatant samples in duplicates and interpolated from the VEGF standard curve. The interpolated values are plotted (mean +/- SD, n=3). The mean VEGF concentration was determined to be 34.76 pg/mL in 2 Day PHA-M stimulated PBMC cell culture supernatant, 393.7 pg/mL in 5 Day PHA-M stimulated PBMC cell culture supernatant, and undetectable in unstimulated PBMC cell culture supernatant at both 2 and 5 days (not shown).

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)
  • sELISA

Supplier Data

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)

Serum from ten individual healthy human female donors was measured in duplicate.

Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean VEGF concentration was determined to be 235.0 pg/mL with a range of 81.10 – 993.3 pg/mL.

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)
  • sELISA

Supplier Data

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)

Interpolated concentrations of native VEGF in human breast milk (de-fatted), saliva, and urine samples.

The concentrations of VEGF were measured in duplicates, interpolated from the VEGF standard curves and corrected for sample dilution. Undiluted samples are as follows : breast milk 2%, saliva 12.5%, and urine 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean VEGF concentration was determined to be 21487 pg/mL in neat human breast milk (de-fatted), 3712 pg/mL in neat human saliva, and 262.0 pg/mL in neat human urine.

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)
  • sELISA

Supplier Data

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)

Interpolated concentrations of native VEGF in human serum and plasma samples.

The concentrations of VEGF were measured in duplicates, interpolated from the VEGF standard curves and corrected for sample dilution. Undiluted samples are as follows : serum 50%, plasma (citrate) 50%, plasma (heparin) 50% and plasma (EDTA) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean VEGF concentration was determined to be 268.9 pg/mL in neat serum, 123.9 pg/mL in neat plasma (citrate), 133.8 pg/mL in neat plasma (heparin), and 95.76 pg/mL in neat plasma (EDTA).

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)
  • sELISA

Supplier Data

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)

Interpolated concentrations of native VEGF in PC-3 cell extract (1 Day), PC-3 cell extract (2 Day), HepG2 cell extract, A549 cell extract, and MDA-MB-435S cell extract samples based on a 300 μg/mL extract load. The concentrations of VEGF were measured in duplicate and interpolated from the VEGF standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean VEGF concentration was determined to be 86.65 pg/mL in PC-3 cell extract (1 Day), 149.6 pg/mL in PC-3 cell extract (2 Day), 127.7 pg/mL in HepG2 cell extract, 241.7 pg/mL in A549 cell extract, and 454.5 pg/mL in MDA-MB-435S cell extract.

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)
  • sELISA

Supplier Data

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)

Serial dilutions of recombinant human VEGF189 and VEGF121 were prepared and assayed in parallel with recombinant VEGF165.

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)
  • sELISA

Lab

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)

Standard curve comparison between the Chemiluminescent Human VEGF SimpleStep ELISA and colorimetric Human VEGF SimpleStep ELISA (ab222510) in Sample Diluent NS +1X Enhancer. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)
  • sELISA

Lab

Sandwich ELISA - Human VEGF ELISA Kit, Chemiluminescent (AB326899)

Dynamic range comparison between the Chemiluminescent Human VEGF SimpleStep ELISA, colorimetric Human VEGF SimpleStep ELISA (ab222510), and leading colorimetric competitor. The Chemiluminescent SimpleStep ELISA kit shows increased sensitivity and increased dynamic range.

Key facts

Detection method

Luminescent

Sample types

Cell culture extracts, Cell culture media, Cerebral Spinal Fluid, Citrate plasma, EDTA Plasma, Heparin Plasma, Milk, Saliva, Serum

Reacts with

Human, Monkey, Dog

Assay type

Sandwich

Results type

Quantitative

Sensitivity

= 1.93 pg/mL

Range

5.49 - 4000 pg/mL

Assay time

55m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

Human VEGF ELISA Kit, Chemiluminescent (ab326899) is a rapid single-wash sandwich 55-min ELISA to measure Human VEGF in Cell culture extracts, Cell culture media, Cerebral Spinal Fluid, Citrate plasma, EDTA Plasma, Heparin Plasma, Milk, Saliva, Serum with a sensitivity of 1.93 pg/mL.

How the assay works

Human VEGF ELISA Kit, Chemiluminescent (ab326899) SimpleStep Ignite™ ELISA employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step during a 30 min only incubation, significantly reducing assay time. By utilizing a chemiluminscent detection method this improves the sensitivity and dynamic range when compared to traditional colorimetric ELISAs.

Assay Specificity

Our SimpleStep Ignite™ ELISA kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.
Please refer to our protocol booklet for more details.

Human VEGF (ab326899) protocol summary

1. Mix : add samples / standards to the wells together with the capture and detector antibody cocktail. Incubate 30 min at room temperature
2. Wash
3. Add ChemiHRP development solution - incubate for 2 min
4. Read the results on a plate reader capable of luminescence detection

Precision

[ { "reproducibilityType": "Intra", "sample": "Supernatant", "replicates": 5, "mean": null, "standardDeviation": null, "coefficientOfVariability": "= 5.4" }, { "reproducibilityType": "Inter", "sample": "Supernatant", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "= 5.5" } ]

Recovery

[ { "sample": "Cell culture extracts", "range": "80 - 94 %", "average": "= 94" }, { "sample": "Cell culture media", "range": "81 - 89 %", "average": "= 89" }, { "sample": "Cerebral Spinal Fluid", "range": "89 - 95 %", "average": "= 95" }, { "sample": "Citrate plasma", "range": "95 - 107 %", "average": "= 107" }, { "sample": "EDTA Plasma", "range": "95 - 99 %", "average": "= 99" }, { "sample": "Heparin Plasma", "range": "84 - 92 %", "average": "= 92" }, { "sample": "Milk", "range": "91 - 98 %", "average": "= 98" }, { "sample": "Saliva", "range": "105 - 118 %", "average": "= 118" }, { "sample": "Serum", "range": "98 - 116 %", "average": "= 116" } ]

What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C

Product protocols

Target data

N-VEGF. Participates in the induction of key genes involved in the response to hypoxia and in the induction of angiogenesis such as HIF1A (PubMed : 35455969). Involved in protecting cells from hypoxia-mediated cell death (By similarity).. VEGFA. Growth factor active in angiogenesis, vasculogenesis and endothelial cell growth (PubMed : 34530889). Induces endothelial cell proliferation, promotes cell migration, inhibits apoptosis and induces permeabilization of blood vessels. Binds to the FLT1/VEGFR1 and KDR/VEGFR2 receptors, heparan sulfate and heparin. Binds to the NRP1/neuropilin-1 receptor. Binding to NRP1 initiates a signaling pathway needed for motor neuron axon guidance and cell body migration, including for the caudal migration of facial motor neurons from rhombomere 4 to rhombomere 6 during embryonic development (By similarity). Also binds the DEAR/FBXW7-AS1 receptor (PubMed : 17446437).. Isoform VEGF165B. Binds to the KDR receptor but does not activate downstream signaling pathways, does not activate angiogenesis and inhibits tumor growth.
See full target information VEGFA
websiteProtocolBooklet
en

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