Rat vascular cell adhesion protein 1 (VCAM-1/CD106) ELISA kit

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Product Details

Description

The Rat vascular cell adhesion protein 1 (VCAM1/CD106) ELISA Kit allows for the in vitro quantitative determination of VCAM1concentrations in serum, plasma, or tissue homogenates. This kit exclusively recognizes rat VCAM1. The kit has been quality controlled with high sensitivity, strong specificity, good linearity, precision less than 10%, recovery with 85%-104%, and high lot-to-lot consistency. Refer to the product manual for more information.

The detection mechanism of this kit is based on the Sandwich-ELISA technique. VCAM1 in the sample is bound to the anti-rat VCAM1antibody immobilized on the microtiter plate and then sandwiched with a Biotin-labeled VCAM1 antibody. The solution color develops into blue after the ordinal addition of HRP-avidin and TMB. The color development is terminated after adding the stop solution, and the color turns from blue to yellow. The color intensity is positively correlated with VCAM1 content in the sample.

VCAM1, also named CD106, is a glycoprotein expressed mainly in endothelial cells upon activation by extracellular stimuli, including reactive oxygen species and pro-inflammatory cytokines, such as TNFα and IL-1. Under an inflammatory condition, VCAM1 directly interacts with α4β1 integrin expressed on leukocytes, mediating leukocyte adhesion and transendothelial migration across endothelial cells. Recent evidence suggests that VCAM1 is closely related to the progression of various immunological disorders, including rheumatoid arthritis, asthma, transplant rejection, and cancer.

Target Name

vascular cell adhesion protein 1 (VCAM-1/CD106)

Alternative Names

Vcam1 ELISA Kit; Vcam-1 ELISA Kit; Vascular cell adhesion protein 1 ELISA Kit; V-CAM 1 ELISA Kit; VCAM-1 ELISA Kit; CD antigen CD106 ELISA Kit

Abbreviation

VCAM1

Uniprot No.

P29534

Species

Rattus norvegicus (Rat)

Sample Types

serum, plasma, tissue homogenates

Detection Range

15.6 ng/mL-1000 ng/mL

Sensitivity

3.9 ng/mL

Assay Time

1-5h

Sample Volume

50-100ul

Detection Wavelength

450 nm

Research Area

Signal Transduction

Assay Principle

quantitative

Measurement

Sandwich

Precision

Linearity

To assess the linearity of the assay, samples were spiked with high concentrations of rat VCAM-1 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
	Sample	Serum(n=4)
1:1	Average %	89
	Range %	85-94
1:2	Average %	95
	Range %	86-103
1:4	Average %	96
	Range %	82-105
1:8	Average %	98
	Range %	87-105

Recovery

The recovery of rat VCAM-1 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.

Sample Type	Average % Recovery	Range
Serum (n=5)	95	89-104
EDTA plasma (n=4)	89	85-93

Typical Data

These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.

ng/ml	OD1	OD2	Average	Corrected
1000	2.614	2.704	2.659	2.525
500	2.145	2.236	2.191	2.057
250	1.516	1.536	1.526	1.392
125	0.863	0.848	0.856	0.722
62.5	0.571	0.549	0.560	0.426
31.25	0.352	0.347	0.350	0.216
15.6	0.234	0.244	0.239	0.105
0	0.135	0.132	0.134

Materials provided

A micro ELISA plate --The 96-well plate has been pre-coated with an anti-rat VCAM antibody.This dismountable microplate can be divided into 12 x 8 strip plates.
Two vials lyophilized standard --Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
One vial Biotin-labeled VCAM antibody (100 x concentrate) (120 μl/bottle) --Act as the detection antibody.
One vial HRP-avidin (100 x concentrate) (120 μl/bottle) --Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
One vial Biotin-antibody Diluent (15 ml/bottle) --Dilute the high concentration Biotin-antibody to an appropriate working solution.
One vial HRP-avidin Diluent (15 ml/bottle) --Dilute the high concentration HRP-avidin solution to an appropriate solution.
One vial Sample Diluent (50 ml/bottle)--Dilute the sample to an appropriate concentration.
One vial Wash Buffer (25 x concentrate) (20 ml/bottle) --- Wash away unbound or free substances.
One vial TMB Substrate (10 ml/bottle) --Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
One vial Stop Solution (10 ml/bottle) --Stop the color reaction. The solution color immediately turns from blue to yellow.
Four Adhesive Strips (For 96 wells) --Cover the microplate when incubation.
An instruction manual

Materials not provided

A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
An incubator can provide stable incubation conditions up to 37°C±5°C.
Centrifuge
Vortex
Squirt bottle, manifold dispenser, or automated microplate washer
Absorbent paper for blotting the microtiter plate
50-300ul multi-channel micropipette
Pipette tips
Single-channel micropipette with different ranges
100ml and 500ml graduated cylinders
Deionized or distilled water
Timer
Test tubes for dilution

ELISA Data Analysis

Watch ELISA data processing video & download Curve Expert if needed

Troubleshooting
and FAQs

ELISA kit FAQs

Storage

Store at 2-8°C. Please refer to protocol.

Lead Time

3-5 working days

Target Background

Function (From Uniprot)	Important in cell-cell recognition. Appears to function in leukocyte-endothelial cell adhesion. Interacts with integrin alpha-4/beta-1 (ITGA4/ITGB1) on leukocytes, and mediates both adhesion and signal transduction. The VCAM1/ITGA4/ITGB1 interaction may play a pathophysiologic role both in immune responses and in leukocyte emigration to sites of inflammation.
Gene References into Functions	Western blotting analyses validated the fact that miR-142-5p inhibited the expression of VCAM-1 protein. Bone marrow-derived mesenchymal stem cells (BMMSCs) transfected with miR-142-5p showed a significantly decreased migration ability in a Transwell migration assay. PMID: 29789783 Panax notoginseng saponins and Ginsenoside protect aorta form atherosclerosis via Rb1 via activation of Nrf2 and suppression of p38 - VCAM-1 signal pathway. PMID: 27620662 Berberine exhibited renoprotective effects in diabetic nephropathy (DN), and the underlying molecular mechanisms may be associated with changes in the levels and regulation of beta-arrestin 1/2 expression, as well as intercellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) levels in the kidney. PMID: 26531813 The combination of DC vaccination and antiangionesis inhibited the rats with malignant glioma by stimulating immune response, supressing the expression of angiogenesis-related proteins VEGF, VCAM, and ICAM-1 PMID: 27072237 VCAM1 was strikingly increased in neurons after intracerebral hemorrhage. PMID: 25868755 Activation of Wnt/beta-catenin signaling in AECs attenuated intercellular adhesion molecule 1 (ICAM-1)/vascular cell adhesion molecule 1 (VCAM-1)-mediated adhesion of both macrophages and neutrophils to AECs. PMID: 26351298 Adjuvant-induced arthritis was not associated with a change in VCAM1 levels. PMID: 25708190 Expression of E-selectin and CAMs was also modified in an age- and ischemia/reperfusion-dependent manner. PMID: 25182537 Upregulation of the adhesion molecules VCAM-1, ICAM-1, and E-selectin. PMID: 24289084 It is reasonable to conclude that PDGFBB significantly enhanced the expression of VCAM-1 in BM-MSCs, which facilitated the migration of BM-MSCs towards PDGFBB. PMID: 24100802 These findings suggested that glioma-induced change in VCAM-1 expression of bone marrow stem cells may play an important role in their tropism towards glioma. PMID: 22562815 Pretreatment of LPS-stimulated vascular endothelial cells with berberine dose-dependently decreased the number of adhered leukocytes and VCAM-1 expression at both RNA and protein levels. PMID: 23058024 results indicate that endothelin-1 stimulates arterial VCAM-1 expression through p38MAPK-dependent activation of neutral sphingomyelinases PMID: 22627111 Clerodendron glandulosum.Coleb leaf extract attenuates in vitro macrophage differentiation and expression of VCAM-1 and P-selectin in thoracic aorta of atherogenic diet fed rats. PMID: 21961520 A cholesterol diet does not play a role in the expression f VCAM-1 in the rat aorta. PMID: 22351665 VCAM1 is expressed at different levels in the various organs of rats after anaphylactic shock. The change of VCAM-1 is the most obvious in lungs. PMID: 21604443 Report specific adhesion and accumulation of VCAM-1-targeted ultrasound microbubbles to inflammatory endothelial cells under hemodynamic shear flow simulation. PMID: 22195873 black tea polyphenol theaflavin suppresses LPS-induced ICAM-1 and VCAM-1 expressions through blockage of NF-kappaB and JNK activation in intestinal epithelial cells PMID: 21184129 The expression of ICAM-1 and VCAM-1 was significantly decreased in the kidney of spontaneously hypertensive rats treated with Cordceps Sinensis. PMID: 20197615 Tanshinone IIA treatment leads to downregulation of ICAM-1 and VCAM-1 protein expression in brain microvascular endothelial cells. PMID: 20687137 Isoflurane, sevoflurane and desflurane down-regulated the expression of VCAM-1 in a dose-dependent manner. PMID: 20936719 Data indicates single-walled carbon nanotube exposure induces oxidative stress, thereby altering ICAM-1 and VCAM-1 expression. PMID: 21121367 Puerarin inhibits the expression of VCAM and P-selectin in the aorta of diabetic rats. PMID: 18095572 Fenofibrate can effectively prevent atherosclerosis by restoring NO concentration and down-regulating VCAM-1 expression. PMID: 18159007 Gene expressions of LOX-1 and VCAM-1 were higher in the aortic arch of HIV-1transgenic rats (Tg); the level of sICAM-1 was elevated in the HIV-1Tg rats but the ICAM-1 gene expression profile did not show any differences. PMID: 19997643 VCAM-1(+) immunoreactivity indicate an important role of choroid plexus in the induction of acute experimental autoimmune encephalomyelitis PMID: 19922364 Integrin alpha2beta1 on rat myeloma cells modulates interaction of alpha4beta1 integrin with vascular cell adhesion molecule-1 but not fibronectin. PMID: 12160302 During experimental allergic encephalomyelitis, a strong activation of spinal perivascular macrophages is observed accompanied by a consistently increased expression of VCAM-1. PMID: 12196270 iNOS and VCAM-1 are induced by angiotensin II in a pathway that involves p38 MAPK PMID: 15001568 Results are consistent with a role for the integrin alpha4beta1--vascular cell adhesion molecule-1 interaction in extension of sympathetic axons into the myocardium. PMID: 15366013 Suppression of the expression levels of VCAM-1 is associated with the amelioration of chronic rejection by various drugs, suggesting that these molecules are important key molecules in chronic rejection. PMID: 15919506 clusters of VCAM-1(+) immunoreactive cells localized in the choroid plexus epithelium and hypothalamo-hypophyses portal vessel system indicate an inflammation in acute autoimmune encephalitis PMID: 17064783 These results suggest that P2Y2R activation in salivary gland cells increases the EGFR-dependent expression of VCAM-1 and the binding of lymphocytes. PMID: 17599409 candidate genes for salt-sensitive hypertension in the SHRSP, including Edg1 and Vcam1, which are differentially expressed and lie on common and functionally important pathways. PMID: 17938382 Thromboxane receptor upregulates IL-1beta-induced vascular cell adhesion molecule 1 (VCAM-1) expression by enhancing the activation of c-Jun N-terminal kinase (JNK) pathway PMID: 18032781 the expression of VCAM-1 protein was upregulated in cerebral and common carotid arteries of simulated microgravity rats, and the upregulation of VCAM-1 may contribute to impaired endothelium-dependent relaxation in simulated microgravity rat vasculature PMID: 18221587 VCAM-1 siRNA reduces neointimal formation after surgical mechanical injury of the rat carotid artery. PMID: 19958991 Show More Hide All
Subcellular Location	Membrane; Single-pass type I membrane protein.
Tissue Specificity	Expressed in aortic endothelial cells, with low expression in the descending thoracic aorta and the outer curvature of the aortic arch, where pulsatory shear stress exists, and high in the inner curvature of the aortic arch, where oscillatory shear stress
Database Links	KEGG: rno:25361 STRING: 10116.ENSRNOP00000019377 UniGene: Rn.11267

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Check Code：

Code	CSB-E07275r
Size	96T,5×96T,10×96T
Trial Size
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Intra-assay Precision (Precision within an assay): CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess.

Inter-assay Precision (Precision between assays): CV%<10%

Three samples of known concentration were tested in twenty assays to assess.