CD133 Antibody - #AF5120

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Product: CD133 Antibody
Catalog: AF5120
Description: Rabbit polyclonal antibody to CD133
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Mol.Wt.: 97 kDa; 97kD(Calculated).
Uniprot: O43490
RRID: AB_2837606

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Related Downloads
MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Clonality:
Polyclonal
Specificity:
CD133 Antibody detects endogenous levels of total CD133.
RRID:
AB_2837606
Cite Format: Affinity Biosciences Cat# AF5120, RRID:AB_2837606.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

AC133; Antigen AC133; CD133; CORD12; Hematopoietic stem cell antigen; hProminin; MCDR2; MSTP061; OTTHUMP00000217744; OTTHUMP00000217745; OTTHUMP00000217746; PROM1; PROM1_HUMAN; Prominin I; Prominin like 1; Prominin like protein 1 precursor; Prominin mouse like 1; Prominin-1; Prominin-like protein 1; Prominin1; PROML1; RP41; STGD4;

Immunogens

Immunogen:
Uniprot:

O43490(PROM1_HUMAN) >>Visit HPA database.

Gene(ID):
PROM1(8842)
Expression:
O43490 PROM1_HUMAN:

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).

Description:
CD133, originally known as AC133. CD133 is a glycoprotein also known in humans and rodents as Prominin 1 (PROM1). Currently the function of CD133 is unknown. It is a member of pentaspan transmembrane glycoproteins (5-transmembrane, 5-TM), which specifically localize to cellular protrusions. CD133 is expressed in hematopoietic stem cells
Sequence:
MALVLGSLLLLGLCGNSFSGGQPSSTDAPKAWNYELPATNYETQDSHKAGPIGILFELVHIFLYVVQPRDFPEDTLRKFLQKAYESKIDYDKPETVILGLKIVYYEAGIILCCVLGLLFIILMPLVGYFFCMCRCCNKCGGEMHQRQKENGPFLRKCFAISLLVICIIISIGIFYGFVANHQVRTRIKRSRKLADSNFKDLRTLLNETPEQIKYILAQYNTTKDKAFTDLNSINSVLGGGILDRLRPNIIPVLDEIKSMATAIKETKEALENMNSTLKSLHQQSTQLSSSLTSVKTSLRSSLNDPLCLVHPSSETCNSIRLSLSQLNSNPELRQLPPVDAELDNVNNVLRTDLDGLVQQGYQSLNDIPDRVQRQTTTVVAGIKRVLNSIGSDIDNVTQRLPIQDILSAFSVYVNNTESYIHRNLPTLEEYDSYWWLGGLVICSLLTLIVIFYYLGLLCGVCGYDRHATPTTRGCVSNTGGVFLMVGVGLSFLFCWILMIIVVLTFVFGANVEKLICEPYTSKELFRVLDTPYLLNEDWEYYLSGKLFNKSKMKLTFEQVYSDCKKNRGTYGTLHLQNSFNISEHLNINEHTGSISSELESLKVNLNIFLLGAAGRKNLQDFAACGIDRMNYDSYLAQTGKSPAGVNLLSFAYDLEAKANSLPPGNLRNSLKRDAQTIKTIHQQRVLPIEQSLSTLYQSVKILQRTGNGLLERVTRILASLDFAQNFITNNTSSVIIEETKKYGRTIIGYFEHYLQWIEFSISEKVASCKPVATALDTAVDVFLCSYIIDPLNLFWFGIGKATVFLLPALIFAVKLAKYYRRMDSEDVYDDVETIPMKNMENGNNGYHKDHVYGIHNPVMTSPSQH

PTMs - O43490 As Substrate

Site PTM Type Enzyme
Y34 Phosphorylation
K82 Acetylation
Y219 Phosphorylation
K225 Acetylation
K257 Acetylation
K264 Acetylation
T266 Phosphorylation
S275 Phosphorylation
T276 Phosphorylation
S719 Phosphorylation
S733 Phosphorylation
T739 Phosphorylation
S824 Phosphorylation
Y828 Phosphorylation P12931 (SRC) , P06241 (FYN)
T833 Phosphorylation
Y852 Phosphorylation P12931 (SRC) , P06241 (FYN)
S863 Phosphorylation

Research Backgrounds

Function:

May play a role in cell differentiation, proliferation and apoptosis. 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.

PTMs:

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.

Subcellular Location:

Apical cell membrane>Multi-pass membrane protein. Cell projection>Microvillus membrane>Multi-pass membrane protein. Cell projection>Cilium>Photoreceptor outer segment. Endoplasmic reticulum. Endoplasmic reticulum-Golgi intermediate compartment.
Note: Found in extracellular membrane particles in various body fluids such as cerebrospinal fluid, saliva, seminal fluid and urine.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
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).

Subunit Structure:

Interacts with CDHR1 and with actin filaments. Interacts with NAT8 and NAT8B.

Family&Domains:

Belongs to the prominin family.

Research Fields

· Human Diseases > Cancers: Overview > Transcriptional misregulation in cancer.

References

1). Characteristics of expression of cancer stem cell markers in various approaches to cervical cancer treatment.. Journal of Clinical Oncology [IF=45.3]
2). HYD-PEP06 suppresses hepatocellular carcinoma metastasis, epithelial–mesenchymal transition and cancer stem cell-like properties by inhibiting PI3K/AKT and WNT/β-catenin signaling activation. Acta Pharmaceutica Sinica B (PubMed: 34221870) [IF=14.5]

Application: WB    Species: Human    Sample: HepG2 cells

Figure 5 Tumor spheres isolated from hepatocellular carcinoma HepG2 cells obtained the characteristics of cancer stem cells. (A) Comparison of the protein levels of CD133 and CD44 in HCCLM3 and HepG2 cells. The protein expression of (B) CD133 and (C) CD44 was dramatically increased in HepG2 cells compared with those in HCCLM3 cells. * P<0.05, compared with the HCCLM3 cells. (D) The images of 3D tumor spheroids were captured using the fluorescence microscope every three days of Day 0 in attachment surface plates and Days 0, 3, 5, 7, and 10 in ultra-low attachment surface plates with CSC medium (magnification 200×); scale bar: 20 μm. (E) The protein expression of CD133 and CD44 was analyzed by Western blot in HepG2 and CSC-like cells. The protein expression of (F) CD133 and (G) CD44 was dramatically increased in CSC-like cells compared with HepG2 cells. Data are presented as mean±SEM; * P<0.05, ** P<0.01. (H) The expression of CD133 and CD44 were assessed by immunofluorescence staining in HepG2 and tumor sphere cells. Blue represents DAPI; green represents CD44; red represents CD133. Magnification: 400×. (I) CSC augments tumor-initiating in vivo. Representative xenograft tumors derived from CSC-like cells compared to HepG2 cells after two weeks subcutaneous injection in nude mice (n=3). Left panel: HepG2 cells; right: CSCs. (J) The protein levels of CD133 and CD44 were analyzed by Western blot in the tumor tissues isolated from xenograft models. GAPDH served as a sample loading control. The protein levels of CSC markers (K) CD133 and (L) CD44 were dramatically upregulated in right tissues compared with left tissues. Data are presented as mean±SEM; * P < 0.05, *** P< 0.001.

Application: IF/ICC    Species: Human    Sample: HepG2 cells

Figure 5 Tumor spheres isolated from hepatocellular carcinoma HepG2 cells obtained the characteristics of cancer stem cells. (A) Comparison of the protein levels of CD133 and CD44 in HCCLM3 and HepG2 cells. The protein expression of (B) CD133 and (C) CD44 was dramatically increased in HepG2 cells compared with those in HCCLM3 cells. * P<0.05, compared with the HCCLM3 cells. (D) The images of 3D tumor spheroids were captured using the fluorescence microscope every three days of Day 0 in attachment surface plates and Days 0, 3, 5, 7, and 10 in ultra-low attachment surface plates with CSC medium (magnification 200×); scale bar: 20 μm. (E) The protein expression of CD133 and CD44 was analyzed by Western blot in HepG2 and CSC-like cells. The protein expression of (F) CD133 and (G) CD44 was dramatically increased in CSC-like cells compared with HepG2 cells. Data are presented as mean±SEM; * P<0.05, ** P<0.01. (H) The expression of CD133 and CD44 were assessed by immunofluorescence staining in HepG2 and tumor sphere cells. Blue represents DAPI; green represents CD44; red represents CD133. Magnification: 400×. (I) CSC augments tumor-initiating in vivo. Representative xenograft tumors derived from CSC-like cells compared to HepG2 cells after two weeks subcutaneous injection in nude mice (n=3). Left panel: HepG2 cells; right: CSCs. (J) The protein levels of CD133 and CD44 were analyzed by Western blot in the tumor tissues isolated from xenograft models. GAPDH served as a sample loading control. The protein levels of CSC markers (K) CD133 and (L) CD44 were dramatically upregulated in right tissues compared with left tissues. Data are presented as mean±SEM; * P < 0.05, *** P< 0.001.
3). TGFBI secreted by tumor-associated macrophages promotes glioblastoma stem cell-driven tumor growth via integrin αvβ5-Src-Stat3 signaling. Theranostics (PubMed: 35673564) [IF=12.4]

Application: IF/ICC    Species: Human    Sample:

TGFBI mediates the tumor-promoting effect of M2-like TAMs. (A) Representative immunofluorescence (IF) staining of TGFBI and stem cell markers (CD133, Olig2) in human GBM were shown. Nuclei were counterstained with DAPI (blue). Scale bar represents 20 µm. (B) Representative tumorsphere images of GSCs (456 and 3691) cultured with rhTGFBI or control for 48 hours. Scale Bar: 400 µm. (C) Quantification of tumorspheres in (B). N = 3, **p < 0.01, Student's t-test. (D, E) Cell viability (D) and in vitro limiting dilution (E) assay of GSCs (456 and 3691) cultured with rhTGFBI or control. N = 6 biological independent samples. Data were represented as means ± SD, ****p < 0.0001, Student's t-test (D); *p < 0.05, likelihood ratio test (E). (F) Representative IF images of EdU incorporation. Scale bar represents 20 µm. (G) The quantification of the percentage of EdU+ cells. N = 3, *p < 0.05, **p < 0.01, Student's t-test. (H) IB analysis showed the extent of TGFBI knockdown in M2 macrophages. GAPDH was used as a loading control. (I) Representative HE staining of mouse brain (cross-section) 25 days after transplantation. Scale bar represents one cm. (J) Representative IF images of Ki67 in mouse models. Scale bar represents 200 µm. (K) Kaplan-Meier survival plot of mice co-implanted with GSCs (456) and shTGFBI M2 macrophages or shNT M2 macrophages. N = 9 for each group, **p < 0.01, ***p < 0.001, log-rank test.
4). Polysaccharide from Flammulina velutipes residues protects mice from Pb poisoning by activating Akt/GSK3β/Nrf-2/HO-1 signaling pathway and modulating gut microbiota. International Journal of Biological Macromolecules (PubMed: 36610568) [IF=8.2]
5). Worse treatment response to neoadjuvant chemoradiotherapy in young patients with locally advanced rectal cancer. BMC Cancer (PubMed: 32891131) [IF=3.8]

Application: IHC    Species: Human    Sample:

Fig. 3 CD133 expression was associated with age and prognosis of LARC patients. (a) and (b) Cut-off points for CD133 expression determined by the X-tile program. X-tile analysis divided the entire cohort into the training (shown in the upper-left quartile of A) and matched validation sets (shown on the bottom X-axis of A) based on patient survival data. The black dot in the validation set represents the exact cut-off value for CD133 expression. The entire cohort was divided into low (blue) and high (gray) NLR count groups based on the optimal cut-point, as is shown on a histogram of the entire cohort (b). (c) Kaplan–Meier curve of disease-free survival and (d) overall survival for the optimal cut-point of the CD133 expression. Representative figures of expression of CD133 in rectal cancer tissues (10 × 40), (e) Negative (−), (f) Weakly positive (+), (g) Positive (++), (h) Strongly positive (+++). (i) The CD133 expression scores in the young and old groups. (j) The correlation analysis of CD133 expression score and patient age. (k) The CD133 expression scores in the pCR and non-pCR groups. (l) Kaplan–Meier curve of disease-free survival and (m) overall survival for the young and old group

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