Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen mouse liver tissue labeling ab252933 with VSIG4 at 1/100 dilution followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000) dilution. Positive staining on Kupffer cells of mouse liver (PMID: 17016562) is observed. The nuclear counterstain is DAPI (Blue).

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling VSIG4 with ab252933 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on Kupffer cells of human liver (PMID: 17016562) is observed. Counterstained with Hematoxylin.

Heat mediated antigen retrieval using ab208572 retrieval buffer.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) .

VSIG4 was immunoprecipitated from 0.35 mg of human placenta lysate with ab252933 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab252933 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/1000 dilution.

Lane 1: Human placenta lysate 10 μg (Input).
Lane 2: ab252933 IP in human placenta lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab252933 in human placenta lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.

The molecular weight observed is consistent with what has been described in previous literature (PMID: 16009265, 17016562).

Immunohistochemical analysis of paraffin-embedded human lung tissue labeling VSIG4 with ab252933 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining on macrophages of human lung (PMID: 17016562) is observed. Counterstained with Hematoxylin.

Heat mediated antigen retrieval using ab208572 retrieval buffer.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling VSIG4 with ab252933 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on stroma of human placenta (PMID: 17016562) is observed. Counterstained with Hematoxylin.

Heat mediated antigen retrieval using ab208572 retrieval buffer.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling VSIG4 with ab252933 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on Kupffer cells of mouse liver (PMID: 17016562) is observed. Counterstained with Hematoxylin.

Heat mediated antigen retrieval using ab208572 retrieval buffer.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

All lanes : Anti-VSIG4 antibody [EPR22576-70] (ab252933) at 1/1000 dilution

Lane 1 : Human liver tissue lysate
Lane 2 : Human placenta tissue lysate
Lane 3 : Jurkat (human T cell leukemia cell line from peripheral blood), whole cell lysate
Lane 4 : THP-1 (human monocytic leukemia cell line), whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
Lanes 1-2 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Lanes 3-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 44 kDa
Observed band size: 37,50 kDa why is the actual band size different from the predicted?

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

Exposure time: 125 seconds.

The molecular weight observed is consistent with what has been described in previous literatures (PMID: 16009265, 17016562)

The expression profile observed is consistent with the literatures (PMID: 16009265, 17016562).

Negative control: Jurkat (PMID: 16009265).

This blot was developed using a higher sensitivity ECL substrate.

Anti-VSIG4 antibody [EPR22576-70] (ab252933) at 1/1000 dilution + Mouse liver tissue lysate at 20 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 44 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?

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

Exposure time: 3 minutes.

The expression profile and molecular weight observed is consistent with what has been described in the literature (PMID: 17016562, PMID: 17016562).

This blot was developed using a higher sensitivity ECL substrate.

Anti-VSIG4 antibody [EPR22576-70] (ab252933) at 1/1000 dilution + His-tagged human VSIG4 recombinant protein (aa 19-232) 10 ng

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 44 kDa
Observed band size: 27 kDa why is the actual band size different from the predicted?

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

Exposure time: 3 minutes.