Recombinant
RabMAb

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

Overview

  • Product name

    Anti-CD105 antibody [EPR22811-18]
    See all CD105 primary antibodies
  • Description

    Rabbit monoclonal [EPR22811-18] to CD105
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, ICC/IF, IP, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human CD105 aa 1-600. The exact sequence is proprietary.
    Database link: P17813

  • Positive control

    • WB: HUVEC and U937 whole cell lysate; Human lung and placenta tissue lysate. IHC-P: Human ovarian carcinoma and placenta tissue. ICC/IF: U937 cells. Flow Cyt: HUVEC and U937 cells. IP: HUVEC whole cell lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab231774 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt 1/500.
ICC/IF 1/500.
IP 1/30.
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Predicted molecular weight: 70 kDa.

Target

  • Function

    Major glycoprotein of vascular endothelium. May play a critical role in the binding of endothelial cells to integrins and/or other RGD receptors.
  • Tissue specificity

    Endoglin is restricted to endothelial cells in all tissues except bone marrow.
  • Involvement in disease

    Defects in ENG are the cause of hereditary hemorrhagic telangiectasia type 1 (HHT1) [MIM:187300, 108010]; also known as Osler-Rendu-Weber syndrome 1 (ORW1). HHT1 is an autosomal dominant multisystemic vascular dysplasia, characterized by recurrent epistaxis, muco-cutaneous telangiectases, gastro-intestinal hemorrhage, and pulmonary (PAVM), cerebral (CAVM) and hepatic arteriovenous malformations; all secondary manifestations of the underlying vascular dysplasia. Although the first symptom of HHT1 in children is generally nose bleed, there is an important clinical heterogeneity.
  • Cellular localization

    Membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • AI528660 antibody
    • AI662476 antibody
    • CD 105 antibody
    • CD105 antibody
    • CD105 antigen antibody
    • EGLN_HUMAN antibody
    • END antibody
    • Endoglin antibody
    • Eng antibody
    • FLJ41744 antibody
    • HHT1 antibody
    • ORW antibody
    • ORW1 antibody
    • Osler Rendu Weber syndrome 1 antibody
    • RP11 228B15.2 antibody
    • S endoglin antibody
    • S-endoglin antibody
    • SN6 antibody
    see all

Images

  • Anti-CD105 antibody [EPR22811-18] (ab231774) at 1/1000 dilution + U937 (human histiocytic lymphoma monocyte), whole cell lysate 40 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 70 kDa
    Observed band size: 97 kDa
    why is the actual band size different from the predicted?


    Exposure time: 103 seconds


    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 : 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 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lanes 4-5 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

    Predicted band size: 70 kDa



    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).

  • 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.

  • 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).

  • 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).

  • 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 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. 

  • 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.

References

ab231774 has not yet been referenced specifically in any publications.

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