Anti-CD105 antibody [EPR19911-220]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD105 antibody [EPR19911-220] (AB252345)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling CD105 with ab252345 at 1/2000 dilution, followed by the Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) ready to use. Positive staining on glomerular and peritubular microvasculature of human kidney (PMID : 25381426) is observed. Counter stained with hematoxylin.

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

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.

The section was incubated with ab252345 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD105 antibody [EPR19911-220] (AB252345)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling CD105 with ab252345 at 1/2000 dilution, followed by the Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) ready to use. Positive staining on sinusoidal endothelial cells of human liver (PMID : 30563158) is observed. Counter stained with hematoxylin.

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

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.

The section was incubated with ab252345 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD105 antibody [EPR19911-220] (AB252345)

Immunohistochemical analysis of human breast carcinoma tissue staining CD105 with ab252345 at 1/2000 dilution and a ready to use secondary Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on endothelial cells of human breast carcinoma. Counterstaining was with Hematoxylin. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. The section was incubated with ab252345 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument

• IP

Unknown

Immunoprecipitation - Anti-CD105 antibody [EPR19911-220] (AB252345)

CD105 was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab252345 at 1/20 dilution. Western blot was performed from the immunoprecipitate using ab252345 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : HeLa whole cell lysate 10 μg (Input).
Lane 2 : ab252345 IP in HeLa whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab252345 in HeLa whole cell lysate.

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

All lanes:

Immunoprecipitation - Anti-CD105 antibody [EPR19911-220] (ab252345)

Predicted band size: 118 kDa,70 kDa,85 kDa

Observed band size: 95 kDa

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• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD105 antibody [EPR19911-220] (AB252345)

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling CD105 with ab252345 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) ready to use. Positive staining on peritubular microvasculature of rat kidney (PMID : 25381426) is observed. Counter stained with hematoxylin.

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

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.

The section was incubated with ab252345 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• WB

Lab

Western blot - Anti-CD105 antibody [EPR19911-220] (AB252345)

Lanes 1 - 4 : Merged signal (red and green). Green - ab252345 observed at 95 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab252345 was shown to recognize ENG (Endoglin) in wild-type HeLa cells as signal was lost at the expected MW in ENG knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and ENG knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab252345 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD105 antibody [EPR19911-220] (ab252345) at 1 µg/mL

Lane 1:

Wild-type HeLa whole cell lysate at 20 µg

Lane 2:

ENG knockout HeLa whole cell lysate at 20 µg

Lane 3:

HUVEC whole cell lysate at 20 µg

Predicted band size: 70 kDa

Observed band size: 95 kDa

false

• WB

Unknown

Western blot - Anti-CD105 antibody [EPR19911-220] (AB252345)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times.

Lane 1 : 26 seconds.

Lane 2 : 6 seconds.

Lane 3 : 3 minutes.

The expression profile / molecular weight observed is consistent with what has been described in the literature (PMID : 12746487; 9872992).

All lanes:

Western blot - Anti-CD105 antibody [EPR19911-220] (ab252345) at 1/1000 dilution

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

Rat spleen 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: 70 kDa

Observed band size: 190 kDa,95 kDa

false