Anti-LYVE1 antibody [EPR21771]

• Flow Cyt

Lab

Flow Cytometry - Anti-LYVE1 antibody [EPR21771] (AB218535)

Flow cytometry overlay histogram showing left, bEND.3 treated with 100ng/mL TNF-alpha for 24h and right, negative untreated bEND.3 stained with ab218535 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interactionfollowed by the antibody (ab218535) (1x 106 in 100μl at 10.0 μg/ml (1/209)) for 30min on ice.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice

Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LYVE1 antibody [EPR21771] (AB218535)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling LYVE1 with ab218535 at 1/5000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on the endothelial surface of mouse hepatic sinusoids (PMID : 16353487; PMID : 11719431). Counter stained with Hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-LYVE1 antibody [EPR21771] (AB218535)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver tissue labeling LYVE1 with ab218535 at 1/500 dilution (green), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive staining of the endothelium of sinusoid blood vessels on mouse liver tissue section (PMID : 11719431).

The nuclear counter stain is DAPI (blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• Flow Cyt

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Flow Cytometry - Anti-LYVE1 antibody [EPR21771] (AB218535)

Flow cytometric analysis of bEnd.3 (mouse brain endothelioma cell line) cells labeling LYVE1 with ab218535 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

Gated on total viable cells.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-LYVE1 antibody [EPR21771] (AB218535)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse stomach tissue labeling LYVE1 with ab218535 at 1/5000 dilution (green), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive staining of the endothelium of lymph vessels in the submucosae on mouse stomach tissue section (PMID : 15705793).

The nuclear counter stain is DAPI (blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LYVE1 antibody [EPR21771] (AB218535)

Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling LYVE1 with ab218535 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on the lymphatic endothelial cells of mouse lung is observed. Counter stained with hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LYVE1 antibody [EPR21771] (AB218535)

Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling LYVE1 with ab218535 at 1/5000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on the lymphatic endothelium of mouse colon (PMID : 14722766). Counter stained with Hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

• IP

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Immunoprecipitation - Anti-LYVE1 antibody [EPR21771] (AB218535)

LYVE1 was immunoprecipitated from 0.35 mg mouse lung lysate with ab218535 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab218535 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : Mouse lung lysate 10 μg (Input).

Lane 2 : ab218535 IP in mouse lung lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab218535 in mouse lung lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 8 seconds.

Several bands are observed including soluble, glycosylated and non-glycosylated forms which are consistent with the literature (PMID : 26966180).

All lanes:

Immunoprecipitation - Anti-LYVE1 antibody [EPR21771] (ab218535)

Predicted band size: 35 kDa

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• WB

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Western blot - Anti-LYVE1 antibody [EPR21771] (AB218535)

Exposure time : Lane 1 : 3 minutes; Lane 2 : 32 seconds; Lane 3 : 5 seconds.

Blocking/Dilution buffer : 5% NFDM/TBST.

Several bands are observed including soluble, glycosylated and non-glycosylated forms which are consistent with the literature (PMID : 26966180).

All lanes:

Western blot - Anti-LYVE1 antibody [EPR21771] (ab218535) at 1/1000 dilution

Lane 1:

Mouse lymph node lysate at 20 µg

Lane 2:

bEnd.3 (mouse brain endothelioma cell line) whole cell lysate at 20 µg

Lane 3:

Mouse lung lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Predicted band size: 35 kDa

Observed band size: 34-70 kDa

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