Anti-LYVE1 antibody [RM1008]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LYVE1 antibody [RM1008] (AB281587)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labelling LYVE1 with ab281587 at 1/2000 dilution (0.293 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on endothelial vessels of mouse spleen. The section was incubated with ab281587 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt

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Flow Cytometry - Anti-LYVE1 antibody [RM1008] (AB281587)

Flow cytometric analysis of bEnd.3 (mouse brain endothelioma) cells labelling LYVE1 with 281587 at 1/500 dilution (0.1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730)/Left isotype control. Goat anti rabbit IgG (Alexa Fluor® 647, ab150079) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells. Partial LYVE1 expression on bEnd.3 cells is reported in literature (PMID : 25282873).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LYVE1 antibody [RM1008] (AB281587)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labelling LYVE1 with ab281587 at 1/2000 dilution (0.293 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on blood sinusoids of mouse liver. The section was incubated with ab281587 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-LYVE1 antibody [RM1008] (AB281587)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse lung tissue labeling LYVE1 with ab281587 at 1/2000 dilution (0.228 ug/ml), followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on mouse lung is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using Tris-EDTA buffer (pH9.0) (ab94681).

• IP

Supplier Data

Immunoprecipitation - Anti-LYVE1 antibody [RM1008] (AB281587)

LYVE1 was immunoprecipitated from 0.35 mg Mouse lung lysate with ab281587 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab281587 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse lung lysate 10ug

Lane 2 : ab281587 IP in Mouse lung lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab281587 in Mouse lung lysate

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

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-LYVE1 antibody [RM1008] (ab281587)

Predicted band size: 35 kDa

Observed band size: 50-70 kDa

false

• WB

Lab

Western blot - Anti-LYVE1 antibody [RM1008] (AB281587)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 3 min.

All lanes:

Western blot - Anti-LYVE1 antibody [RM1008] (ab281587) at 1/1000 dilution

All lanes:

A549 (Human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/5000 dilution

Predicted band size: 35 kDa

Observed band size: 50-70 kDa

false

• WB

Lab

Western blot - Anti-LYVE1 antibody [RM1008] (AB281587)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Several bands are observed including soluble, glycosylated and non-glycosylated forms which are consistent with the literature (PMID : 26966180). Exposure time : 37 seconds

All lanes:

Western blot - Anti-LYVE1 antibody [RM1008] (ab281587) at 1/1000 dilution

Lane 1:

Mouse lung lysate at 20 µg

Lane 2:

Mouse brain lysate at 20 µg

Lane 3:

Rat 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 1/100000 dilution

Predicted band size: 35 kDa

Observed band size: 50-70 kDa

false