Recombinant Anti-CD133 antibody [RM1029] (ab284389)

Product name

Anti-CD133 antibody [RM1029]
See all CD133 primary antibodies
Description

Rabbit recombinant multiclonal [RM1029] to CD133
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt, WB, IHC-Fr, IHC-P, ICC, IPmore details
Species reactivity

Reacts with: Mouse, Human
Immunogen

This product was produced with the following immunogens:
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Caco-2, NCCIT, 293T, ES-D3, F9 and Mouse E12.5 brain lysates. IHC-P: Human pancreas, Human kidney, Human breast, Human pancreatic carcinoma, Mouse breast and Mouse liver, Mouse retina tissues. IHC-Fr: and Mouse E14.5 embryonic lung, Mouse E14.5 embryonic kidney tissues. ICC: Caco-2 cells. Flow cyt: Mouse bone marrow and 293T cells. IP: Caco-2, Mouse kidney cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Recombinant Multiclonal
Clone number

RM1029
Isotype

IgG
Research areas
- Neuroscience
- Development

Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab284389 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		1/50.
WB		1/1000. Predicted molecular weight: 97 kDa.
IHC-Fr		1/100. Perform heat mediated antigen retrieval with sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) before commencing with IHC staining protocol.
IHC-P		1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC		1/50. ICC is unsuitable for Mouse
IP		1/30.

Notes
Flow Cyt 1/50.
WB 1/1000. Predicted molecular weight: 97 kDa.
IHC-Fr 1/100. Perform heat mediated antigen retrieval with sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) before commencing with IHC staining protocol.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC 1/50. ICC is unsuitable for Mouse
IP 1/30.

Function

May play a role in cell differentiation, proliferation and apoptosis (PubMed:24556617). Binds cholesterol in cholesterol-containing plasma membrane microdomains and may play a role in the organization of the apical plasma membrane in epithelial cells. During early retinal development acts as a key regulator of disk morphogenesis. Involved in regulation of MAPK and Akt signaling pathways. In neuroblastoma cells suppresses cell differentiation such as neurite outgrowth in a RET-dependent manner (PubMed:20818439).
Tissue specificity

Isoform 1 is selectively expressed on CD34 hematopoietic stem and progenitor cells in adult and fetal bone marrow, fetal liver, cord blood and adult peripheral blood. Isoform 1 is not detected on other blood cells. Isoform 1 is also expressed in a number of non-lymphoid tissues including retina, pancreas, placenta, kidney, liver, lung, brain and heart. Found in saliva within small membrane particles. Isoform 2 is predominantly expressed in fetal liver, skeletal muscle, kidney, and heart as well as adult pancreas, kidney, liver, lung, and placenta. Isoform 2 is highly expressed in fetal liver, low in bone marrow, and barely detectable in peripheral blood. Isoform 2 is expressed on hematopoietic stem cells and in epidermal basal cells (at protein level). Expressed in adult retina by rod and cone photoreceptor cells (at protein level).
Involvement in disease

Retinitis pigmentosa 41
Cone-rod dystrophy 12
Stargardt disease 4
Retinal macular dystrophy 2
Sequence similarities

Belongs to the prominin family.
Post-translational
modifications

Isoform 1 and isoform 2 are glycosylated.
Acetylation at Lys-225, Lys-257 and Lys-264 by NAT8 and NAT8B may control PROM1 protein expression and its function in cell apoptosis.
Cellular localization

Apical cell membrane. Cell projection, microvillus membrane. Cell projection, cilium, photoreceptor outer segment. Endoplasmic reticulum. Endoplasmic reticulum-Golgi intermediate compartment. Found in extracellular membrane particles in various body fluids such as cerebrospinal fluid, saliva, seminal fluid and urine.
Target information above from: UniProt accession O43490 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 8842 Human
- Entrez Gene: 19126 Mouse
- Omim: 604365 Human
- SwissProt: O43490 Human
- SwissProt: O54990 Mouse
- Unigene: 614734 Human
- Unigene: 6250 Mouse
Alternative names
- AC133 antibody
- Antigen AC133 antibody
- CD133 antibody
- CORD12 antibody
- Hematopoietic stem cell antigen antibody
- hProminin antibody
- MCDR2 antibody
- MSTP061 antibody
- OTTHUMP00000217744 antibody
- OTTHUMP00000217745 antibody
- OTTHUMP00000217746 antibody
- PROM1 antibody
- PROM1_HUMAN antibody
- Prominin I antibody
- Prominin like 1 antibody
- Prominin like protein 1 precursor antibody
- Prominin mouse like 1 antibody
- Prominin-1 antibody
- Prominin-like protein 1 antibody
- Prominin1 antibody
- PROML1 antibody
- RP41 antibody
- STGD4 antibody
see all

Western blot - Anti-CD133 antibody [RM1029] (ab284389)

All lanes : Anti-CD133 antibody [RM1029] (ab284389) at 1/1000 dilution

Lane 1 : Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : NCCIT (Human pluripotent embryonic carcinoma epithelial cell) whole cell lysate
Lane 3 : 293T (Human embryonic kidney epithelial cell) whole cell lysate
Lane 4 : ES-D3 (Mouse embryo spherical colony) whole cell lysate
Lane 5 : MEF (Mouse embryonic fibroblast (immortalized) whole cell lysate
Lane 6 : F9 (Mouse embryonal carcinoma epithelial cell) whole cell lysate
Lane 7 : Mouse E12.5 brain lysate
Lane 8 : Rat E12.5 brain lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Predicted band size: 97 kDa

Blocking and diluting buffer and concentration: 5%

NFDM/TBST Negative control: MEF (PMID: 21709693)

Similar band pattern was observed in literature. (PMID: 26137114)

Lysate were not boiled to avoid protein aggregation.

Exposure time: 1 min
Immunocytochemistry - Anti-CD133 antibody [RM1029] (ab284389)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized Caco-2 cells labelling CD133 with ab284389 at 1/50 (10.16 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing membranous staining in Caco-2 cells is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [RM1029] (ab284389)

Immunohistochemical analysis of paraffin-embedded Mouse retina tissue labelling CD133 with ab284389 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on the cone and rod cells of mouse retina. The section was incubated with ab284389 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Flow Cytometry - Anti-CD133 antibody [RM1029] (ab284389)

Flow cytometric analysis of 293T (Human embryonic kidney epithelial cell, Left), Caco-2 (Human colorectal adenocarcinoma epithelial cell, Right) cells labelling CD133 with ab284389 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/2000 dilution was used as the secondary antibody. Negative control: 293T. Gated on viable cells.
Immunohistochemistry (Frozen sections) - Anti-CD133 antibody [RM1029] (ab284389)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse E14.5 embryonic kidney tissue labeling CD133 with ab284389 at 1/100 (5.08 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution (Green). Positive staining on the apical side of mouse E14.5 embryonic kidney buds is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/ml dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Immunoprecipitation - Anti-CD133 antibody [RM1029] (ab284389)

CD133 was immunoprecipitated from 0.35 mg Mouse kidney lysate 10ug with ab284389 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab284389 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Mouse kidney lysate 10ug

Lane 2: abab284389 IP in Mouse kidney lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab284389 in Mouse kidney lysate

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

Exposure time: 5 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [RM1029] (ab284389)

Immunohistochemical analysis of paraffin-embedded Human pancreatic carcinoma tissue labelling CD133 with ab284389 at 1/500 (1.016 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Apical staining on human pancreatic carcinoma. The section was incubated with ab284389 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Flow Cytometry - Anti-CD133 antibody [RM1029] (ab284389)

Flow cytometric analysis of Mouse bone marrow cell cells labelling CD133 with ab284389 at 1/50 dilution (1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/2000 dilution was used as the secondary antibody. Cells were co-stained with anti-CD117 conjugated to APC/Cy7. Gated on viable cells.
Immunoprecipitation - Anti-CD133 antibody [RM1029] (ab284389)

CD133 was immunoprecipitated from 0.35 mg Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate 10ug with ab284389 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab284389 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1: Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate 10ug

Lane 2: ab284389 IP in Caco-2 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab284389 in Caco-2 whole cell lysate

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

Exposure time: 32 seconds.
Flow Cytometry - Anti-CD133 antibody [RM1029] (ab284389)

Flow cytometric analysis of Mouse bone marrow cell cells labelling CD133 with ab284389 at 1/50 dilution (1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/2000 dilution was used as the secondary antibody. Cells were stained with rabbit IgG or ab271092. Then stained with Lineage cocktail conjugated to FITC, anti-Sca-1 conjugated to PE/Cy7, anti-CD117 conjugated to APC/Cy7 and anti-CD16/32 conjugated to BV510.Gated on viable MP cells. Positive staining on CD16/32+ MP cells is consistent with literature. (PMID: 23509298)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [RM1029] (ab284389)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labelling CD133 with ab284389 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on the biliary ducts of mouse liver. The section was incubated with ab284389 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [RM1029] (ab284389)

Immunohistochemical analysis of paraffin-embedded Mouse breast tissue labelling CD133 with ab284389 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Apical staining on mouse breast. The section was incubated with ab284389 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [RM1029] (ab284389)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labelling CD133 with ab284389 at 1/500 (1.016 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Apical staining on human breast. The section was incubated with ab284389 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [RM1029] (ab284389)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling CD133 with ab284389 at 1/500 (1.016 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on parietal layer of Bowman's capsule and apical staining on renal tubules in human kidney. The section was incubated with ab284389 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD133 antibody [RM1029] (ab284389)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labelling CD133 with ab284389 at 1/500 (1.016 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on the pancreatic ducts in human pancreas. The section was incubated with ab284389 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Frozen sections) - Anti-CD133 antibody [RM1029] (ab284389)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse E14.5 embryonic lung tissue labeling CD133 with ab284389 at 1/100 (5.08 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution (Green). Positive staining on the apical side of mouse E14.5 embryonic lung buds is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/ml dilution.

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

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab284389? Please let us know so that we can cite the reference in this datasheet.

ab284389 has been referenced in 1 publication.

Wang Y et al. Overexpression of FOXD2-AS1 enhances proliferation and impairs differentiation of glioma stem cells by activating the NOTCH pathway via TAF-1. J Cell Mol Med 26:2620-2632 (2022). PubMed: 35419917

Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Switch on our new BETA site

Now available

In the coming months

Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Compatible Secondaries

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (1)

Customer reviews and Q&As

Post-translational
modifications