16-0519-81

antibody from Invitrogen Antibodies

Targeting: ITGB3 CD61, GP3A, GPIIIa

Immunohistochemistry (Recommended by provider)

Flow cytometry (Supportive data in Antibodypedia)

Blocking/Neutralizing (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

16-0519-81

Provider

Invitrogen Antibodies

Product name

CD51/CD61 (Integrin alpha v beta 3) Monoclonal Antibody (23C6), Functional Grade, eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The 23C6 monoclonal antibody reacts with the human CD51/CD61 dimer, also known as the integrin alphav/beta3. CD51, an ~120 kDa surface molecule can also non-covalently associate with other beta subunits of the integrin family including beta1 (CD29), beta5 and beta6 to form receptors for extracellular matrix components. Heterodimers of CD51/CD61 are expressed by melanoma cells, endothelial cells and osteoclasts and at very low levels by platelets. The CD51/CD61 complex mediates adhesion to fibrinogen, fibronectin, vitronectin and thrombospondin. Applications Reported: This 23C6 antibody has been reported for use in flow cytometric analysis, and immunohistology staining of frozen tissue sections. 23C6 has also been reported in blocking of some adhesive processes. Applications Tested: This 23C6 antibody has been tested by flow cytometric analysis of human melanoma cell line and peripheral blood. This can be used at less than or equal to 1 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Storage and handling: Use in a sterile environment. Filtration: 0.2 µm post-manufacturing filtered. Purity: Greater than 90%, as determined by SDS-PAGE. Endotoxin Level: Less than 0.001 ng/µg antibody, as determined by LAL assay. Aggregation: Less than 10%, as determined by HPLC.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

23C6

Vial size

50 µg

Concentration

1 mg/mL

Storage

4° C

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Staining of platelets with 0.5 µg of Mouse IgG1 kappa Isotype Control Purified (Product # 14-4714-82) (open histogram), or 0.5 µg of Anti-Human CD51/CD61 (Integrin alpha V beta 3) Purified followed by Anti-Mouse IgG FITC (Product # 11-4011-85) (filled histogram).

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 3 Expression of alphavbeta3 by HUVECs under different conditions ( a ) Flow cytometry analysis demonstrated significant expression of integrin alphavbeta3 in overnight starved cells. (i) Side scatter (SSC) vs. forward scatter (FSC) plot used to gate live cells and resulting (ii) histograms from which alphavbeta3 expression was (iii) quantified for different culture conditions. Colors in (ii) match the conditions shown in the x-axis of (iii). Values were expressed as mean +- SD, n = 3. p < 0.005 was considered significant. ( b ) Immunostaining was performed by (i) staining HUVECs with anti-human CD51/CD61 antibody to detect integrin alphavbeta3 (green). Nuclei were stained by DAPI (blue). Integrin alphavbeta3 expression was highest under overnight starved conditions when compared to control and 2 h starvation. (ii) Quantification of the images in (i) obtained using Image J. Value was expressed as mean +- SD, n = 3. p < 0.05 was considered significant.