Anti-CD105 antibody [EPR22811-18]

• Flow Cyt

Unknown

Flow Cytometry - Anti-CD105 antibody [EPR22811-18] (AB231774)

Flow cytometric analysis of Jurkat (human T cell leukemia T lymphocyte, Left) / HUVEC (human umbilical vein endothelial cell, Right) cells labeling CD105 with ab231774 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control : Jurkat (PMID : 28351936). Gated on viable cells.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD105 antibody [EPR22811-18] (AB231774)

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma tissue labeling CD105 with ab231774 at 1/100 dilution (5.1 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on endothelial cells of human ovarian carcinoma (PMID : 17502949). The section was incubated with ab231774 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

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).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CD105 antibody [EPR22811-18] (AB231774)

Immunofluorescent analysis of 100% methanol-fixed U-937 (human histiocytic lymphoma monocyte) cells labeling CD105 with ab231774 at 1/500 dilution, followed by a ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in U-937 cells is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

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

Negative control : Jurkat (PMID : 28351936).

• Flow Cyt

Unknown

Flow Cytometry - Anti-CD105 antibody [EPR22811-18] (AB231774)

Flow cytometric analysis of U-937 (human histiocytic lymphoma monocyte) cells labeling CD105 with ab231774 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD105 antibody [EPR22811-18] (AB231774)

Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling CD105 with ab231774 at 1/100 dilution (5.1 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human placental trophoblasts (PMID : 17956952) is observed. The section was incubated with ab231774 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

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).

• IP

Unknown

Immunoprecipitation - Anti-CD105 antibody [EPR22811-18] (AB231774)

CD105 was immunoprecipitated from 0.35 mg HUVEC (human umbilical vein endothelial cell) whole cell lysate with ab231774 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab231774 1/1000 dilution (0.51 μg/ml). VeriBlot for IP secondary antibody (HRP) (ab131366) was used as the secondary antibody at 1/5000 dilution.

Lane 1 : HUVEC (human umbilical vein endothelial cell) whole cell lysate 10μg
Lane 2 : ab231774 IP in HUVEC whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab231774 in HUVEC whole cell lysate.

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

Exposure time : 3 mins.

All lanes:

Immunoprecipitation - Anti-CD105 antibody [EPR22811-18] (ab231774)

Predicted band size: 70 kDa

false

• WB

Unknown

Western blot - Anti-CD105 antibody [EPR22811-18] (AB231774)

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

Exposure times : Lanes 1-3 : 26 secs; Lanes 4-5 : 3 mins.

The molecular weight observed is consistent with what has been described in the literature (PMID : 8932339).

This blot was developed using a higher sensitivity ECL substrate.

Negative control : Jurkat (PMID : 28351936); Raji (PMID : 28351936).

All lanes:

Western blot - Anti-CD105 antibody [EPR22811-18] (ab231774) at 1/1000 dilution

Lane 1:

HUVEC (human umbilical vein endothelial cell), whole cell lysate 20 μg

Lane 2:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate 20 μg

Lane 3:

Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate 20 μg

Lane 4:

Human lung tissue lysate 20 μg

Lane 5:

Human placenta tissue lysate 20 μg

Secondary

Lanes 1 - 3:

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

Lanes 4 - 5:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 70 kDa

false

• WB

Unknown

Western blot - Anti-CD105 antibody [EPR22811-18] (AB231774)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 8932339).

All lanes:

Western blot - Anti-CD105 antibody [EPR22811-18] (ab231774) at 1/1000 dilution

All lanes:

U937 (human histiocytic lymphoma monocyte), whole cell lysate 40 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 1/20000 dilution

Predicted band size: 70 kDa

Observed band size: 97 kDa

false

Exposure time: 103s