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<strong>Figure 1 Western Blot Validation in Human Colon Tissue </strong><br>Loading: 15 μg of lysates per lane.Antibodies: TEM-1 4357, (A, 0.5 μg/mL; B, 1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines</strong><br>Loading: 15 μg of lysates per lane.Antibodies: TEM-1 4357, (2 μg/mL), TEM-1 4359, ( 1μg/mL), and GAPDH (0.02 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 Western Blot Validation in Human Cell Lines</strong><br>Loading: 15 μg of lysates per lane.Antibodies: TEM-1 4357, (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Immunohistochemistry Validation of TEM-1 in Human Colon Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded Human Colon tissue using anti-TEM-1 antibody (4357) at 2.5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 5 Immunofluorescence Validation of TEM-1 in Human Colon Tissue</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed Human Colon Tissue labeling TEM-1 with 4357 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
Figure 1 Western Blot Validation in Human Colon Tissue
Loading: 15 μg of lysates per lane.Antibodies: TEM-1 4357, (A, 0.5 μg/mL; B, 1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
IAV
Trial Size

TEM1 Antibody

CATALOG NUMBER: 4357

Clonality:
Polyclonal
Tested Applications:
ELISA, IF, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human
Homology:
Predicted species reactivity based on immunogen sequence: Mouse (86%); Rat: (86%)
Immunogen:
Anti-TEM-1 antibody (4357) was raised against a peptide corresponding to 14 amino acids near the carboxy terminus of human TEM-1.
The immunogen is located within the last 50 amino acids of TEM-1.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IF, IHC-P, WB
Application Note:
WB: 0.5-2 μg/mL; IHC-P: 2.5 μg/mL; IF: 20 μg/mL.
Antibody validated: Western Blot in human and mouse samples; Immunohistochemistry in human samples and Immunofluorescence in human samples. All other applications and species not yet tested.
Specificity:
At least two isoforms of TEM1 are known to exist; this antibody recognizes both isoforms.
Positive Control 1:
Cat. No. 1320 - Human Colon Tissue Lysate
Positive Control 2:
Cat. No. 1223 - Caco-2 Cell Lysate
Positive Control 3:
Cat. No. 1219 – MCF-7 Cell Lysate
Positive Control 4:
Cat. No. 1220 – SK-N-SH Cell Lysate
Predicted Molecular Weight:
Predicted: 81kD and 46kD
Observed: 90 kD (post-modifications: 27 O-linked glycosylations) and 46kD
Validation:

Independent Antibody Validation in Cell lines (Figure 2) shows similar TEM-1 expression profile in human and mouse cell lines detected by two independent anti-TEM-1 antibodies that recognize different epitopes, 4357 against C-terminus domain and 4359 against N-terminus domain.  TEM-1 proteins are detected in the most tested cell lines at different expression levels by the two independent antibodies. 

Isoforms:
Human TEM-1 has 2 isoforms, including isoform 1 (757aa, 81kD) and isoform 2 (433aa, 46kD). Mouse TEM-1 has one isoform (765aa, 82kD) and rat TEM-1 also has one isoform (764aa, 82kD). 4357 can detect all isoforms of human, mouse and rat.
Purification:
TEM1 Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
TEM1 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
TEM1 antibody can be stored at 4°C up to one year. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
CD248
Additional Names:
TEM1 Antibody: TEM1, CD164L1, CD248, Endosialin, Tumor endothelial marker 1
Protein Accession Number:
NP_065137
Protein Gi Number:
9966885
Ncbi Gene Id Number:
57124
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
TEM1 Antibody: Tumor endothelial marker (TEM) 1 was originally identified as a human embryonic fibroblast-specific antigen and was later determined to be endosialin, a single-pass transmembrane glycoprotein that has multiple extracellular domains, including three EGF-like domains, a sushi-like domain, and a C lectin-like domain. TEM proteins are significantly up-regulated during angiogenesis and neoangiogenesis that are crucial for the growth of solid tumors. While TEM1 is not required for angiogenesis during fetal development, postnatal growth or wound healing, it plays a role in tumor growth, invasion, and metastasis. Fibronectin and collagen types I and IV act as specific ligands of TEM1, leading to suggestions that these molecules may cause changes in the extracellular matrix, cell adhesion and migration during tumor invasion.
Background Reference 1:
Rettig et al. Proc. Natl. Acad. Sci. USA 1992; 89:10832-6.
Background Reference 2:
Christian et al. J. Biol. Chem. 2001; 276:7408-14.
Background Reference 3:
Nanda et al. Curr. Opin. Oncol. 2004; 16:44-9.
Background Reference 4:
Nanda et al. Proc. Natl. Acad. Sci. USA 2006; 103:3351-6.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

4357

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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