701257

antibody from Invitrogen Antibodies

Targeting: SELP CD62, CD62P, GMP140, GRMP, PADGEM, PSEL

Western blot (Data presented on provider website)

ELISA (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

701257

Provider

Invitrogen Antibodies

Product name

P-Selectin Recombinant Rabbit Monoclonal Antibody (3H20L10)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with mouse, rat and non-human primate based on sequence homology. Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain. Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

3H20L10

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Carbon nano-onion-mediated dual targeting of P-selectin and P-glycoprotein to overcome cancer drug resistance.

Wang H, Liang Y, Yin Y, Zhang J, Su W, White AM, Bin Jiang, Xu J, Zhang Y, Stewart S, Lu X, He X
Nature communications 2021 Jan 12;12(1):312

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The cancer glycocalyx mediates intravascular adhesion and extravasation during metastatic dissemination.

Offeddu GS, Hajal C, Foley CR, Wan Z, Ibrahim L, Coughlin MF, Kamm RD
Communications biology 2021 Feb 26;4(1):255

ELISA

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Indirect ELISA analysis of P-selectin in Jurkat lysate coated onto the plate using a P-selectin recombinant rabbit monoclonal antibody (Product # 701257) at various dilutions. A non-linear regression analysis was performed (4 PL) and LOD and LOQ for the antibody were determined.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of P-selectin in U2OS cells using a P-selectin recombinant rabbit monoclonal antibody (Product # 701257) followed by detection using an Alexa Fluor 488-conjugated goat anti-rabbit secondary antibody (green) (Image A). Nuclei were stained using DAPI (Image B) and actin stained with Alexa Fluor 594 phalloidin (red) (image C). Image D is a composite image showing membrane localization of P-selectin.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of P-selectin in U2OS cells using a P-selectin recombinant rabbit monoclonal antibody (Product # 701257) followed by detection using an Alexa Fluor 488-conjugated goat anti-rabbit secondary antibody (green) (Image A). Nuclei were stained using DAPI (Image B) and actin stained with Alexa Fluor 594 phalloidin (red) (image C). Image D is a composite image showing membrane localization of P-selectin.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of P-selectin/CD62P was performed on Jurkat cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0. 25% Tritonª X-100 for 20 minutes, and blocked with 5% BSA for 1 hour at room temperature. Cells were labeled with ABfinityª P-selectin/CD62P recombinant rabbit monoclonal antibody (Product # 701257, red histogram) or with rabbit isotype control (pink histogram) at a dilution of 1:250 in 2.5% BSA. After incubation at room temperature for 3 hours, the cells were labeled with Alexa Fluor¨ 488 goat anti-Rabbit Secondary antibody (Product # A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune¨ Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Indirect ELISA analysis of P-selectin in Jurkat lysate coated onto the plate using a P-selectin recombinant rabbit monoclonal antibody (Product # 701257) at various dilutions. A non-linear regression analysis was performed (4 PL) and LOD and LOQ for the antibody were determined.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 5 P-selectin targeting capability of the FSCNO nanoparticles under both static and dynamic/microfluidic cultures. a Confocal images showing the expression of P-selectin in activated human umbilical vein endothelial cells (aHUVECs) but not in HUVECs without activation. b Confocal images of precooled aHUVECs after incubated with SCNO-DH and FSCNO-DH nanoparticles (10 mug mL -1 for DOX and 5 mug mL -1 for HM). The cells were precooled in ice for 3 h to stop its metabolic and uptake activity. c Top and side views of the microfluidic device for studying the P-selectin targeting capability of the FSCNO-DH nanoparticles under dynamic culture condition. Detached cells can be injected into device through inlet2 or inlet3 and cultured in the chambers. Medium containing nanoparticles can be injected through inlet1 and flows out the device via outlet1. Also shown are the computational modeling results of velocity distribution at the middle planes of the channels and chambers, in the top and bottom PDMS parts as indicated by the red dashed lines i and ii, respectively. The data show that the velocity of injected nanoparticle solution (inlet1) is nearly homogeneous in the two chambers. d, e Scheme and confocal images of green fluorescence protein (GFP)-expression HUVECs (in the chamber next to outlet1 for d and in the chamber next to inlet1 for e ) and aHUVECs (in the chamber next to inlet1 for d and in the chamber next to outlet1 for e ) cultured in the microfluidic device for 12 and