Revision 4

#17545Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IHC-P, FC-FP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

75

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q13641

Entrez-Gene Id:

7162

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunohistochemistry (Paraffin) 1:100 - 1:400
Flow Cytometry (Fixed/Permeabilized) 1:50 - 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

TPBG/5T4 (E3M5R) XP® Rabbit mAb recognizes endogenous levels of total TPBG/5T4 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the extracellular domain of human TPBG/5T4 protein.

Background

Trophoblast glycoprotein (TPBG/5T4) is a type I transmembrane glycoprotein that is normally expressed by trophoblast cells of the placenta with a very limited expression pattern in normal adult tissues (1,2). The extracellular domain of TPBG is extensively glycosylated and contains multiple leucine-rich repeats while its cytoplasmic domain consists of a PDZ domain-binding motif, which is important for linking TPBG to intracellular signaling networks involved in the regulation of cell motility and adhesion (3,4). Research studies have shown that cell surface expression of TPBG plays a critical role in modulating signaling cascades that drive cell adhesion, morphology, and motility processes that are fundamental for normal progression of embryogenesis (5-7).

Research studies have demonstrated that TPBG is aberrantly overexpressed in numerous types of solid tumors (8) and functions to promote enhanced tumor cell motility and metastasis (9,10). In some tumors, such as NSCLC and HNSCC, TPBG has been identified as a novel marker of tumor-initiating cells (11,12). The observed differential expression of TPBG by normal tissue versus tumor tissue has been exploited by multiple immunotherapeutic agents that are currently being evaluated for targeting of multiple types of solid tumors (13).

  1. Hole, N. and Stern, P.L. (1990) Int J Cancer 45, 179-84.
  2. Woods, A.M. et al. (2002) Biochem J 366, 353-65.
  3. Shaw, D.M. et al. (2002) Biochem J 363, 137-45.
  4. Awan, A. et al. (2002) Biochem Biophys Res Commun 290, 1030-6.
  5. Barrow, K.M. et al. (2005) Dev Dyn 233, 1535-45.
  6. Ward, C.M. et al. (2003) J Cell Sci 116, 4533-42.
  7. Ward, C.M. et al. (2006) Exp Cell Res 312, 1713-26.
  8. Southall, P.J. et al. (1990) Br J Cancer 61, 89-95.
  9. Carsberg, C.J. et al. (1996) Int J Cancer 68, 84-92.
  10. Myers, K.A. et al. (1994) J Biol Chem 269, 9319-24.
  11. Damelin, M. et al. (2011) Cancer Res 71, 4236-46.
  12. Kerk, S.A. et al. (2017) Clin Cancer Res 23, 2516-2527.
  13. Stern, P.L. and Harrop, R. (2017) Cancer Immunol Immunother 66, 415-426.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IHC-P: Immunohistochemistry (Paraffin) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SignalStain is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 4
#17545

TPBG/5T4 (E3M5R) XP® Rabbit mAb

Western Blotting Image 1: TPBG/5T4 (E3M5R) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from control MCF7 cells (lane 1) or TPBG/5T4 knockout cells (lane 2) using TPBG/5T4 (E3M5R) XP® Rabbit mAb #17545 (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the TPBG/5T4-knockout MCF7 cells confirms specificity of the antibody for TPBG/5T4.
Western Blotting Image 2: TPBG/5T4 (E3M5R) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using TPBG/5T4 (E3M5R) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). Negative expression of TPBG/5T4 protein in Raji cells is consistent with the predicted expression pattern.
Western Blotting Image 3: TPBG/5T4 (E3M5R) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human TPBG/5T4 (hTPBG/5T4-Myc/DDK; +), using TPBG/5T4 (E3M5R) XP® Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunohistochemistry Image 1: TPBG/5T4 (E3M5R) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using TPBG/5T4 (E3M5R) XP® Rabbit mAb.
Immunohistochemistry Image 2: TPBG/5T4 (E3M5R) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human squamous cell carcinoma of the cervix using TPBG/5T4 (E3M5R) XP® Rabbit mAb.
Immunohistochemistry Image 3: TPBG/5T4 (E3M5R) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human esophageal carcinoma using TPBG/5T4 (E3M5R) XP® Rabbit mAb.
Immunohistochemistry Image 4: TPBG/5T4 (E3M5R) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human soft tissue squamous cell carcinoma using TPBG/5T4 (E3M5R) XP® Rabbit mAb.
Immunohistochemistry Image 5: TPBG/5T4 (E3M5R) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human squamous cell carcinoma of the tongue using TPBG/5T4 (E3M5R) XP® Rabbit mAb.
Immunohistochemistry Image 6: TPBG/5T4 (E3M5R) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using TPBG/5T4 (E3M5R) XP® Rabbit mAb.
Immunohistochemistry Image 7: TPBG/5T4 (E3M5R) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human tonsil using TPBG/5T4 (E3M5R) XP® Rabbit mAb.
Immunohistochemistry Image 8: TPBG/5T4 (E3M5R) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human thymus using TPBG/5T4 (E3M5R) XP® Rabbit mAb.
Immunohistochemistry Image 9: TPBG/5T4 (E3M5R) XP® Rabbit mAb Expand Image

Immunohistochemical analysis of paraffin-embedded normal human placenta using TPBG/5T4 (E3M5R) XP® Rabbit mAb.

Immunohistochemistry Image 10: TPBG/5T4 (E3M5R) XP® Rabbit mAb Expand Image

Immunohistochemical analysis of paraffin-embedded normal human brain using TPBG/5T4 (E3M5R) XP® Rabbit mAb.

Immunohistochemistry Image 11: TPBG/5T4 (E3M5R) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using TPBG/5T4 (E3M5R) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemistry Image 12: TPBG/5T4 (E3M5R) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded MCF7 cell pellet (left, positive) or Raji cell pellet (right, negative) using TPBG/5T4 (E3M5R) XP® Rabbit mAb.
Flow Cytometry Image 1: TPBG/5T4 (E3M5R) XP® Rabbit mAb Expand Image
Flow cytometric analysis of fixed, permeabilized Raji cells (blue, negative) and MCF7 cells (green, positive) using TPBG/5T4 (E3M5R) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used a s a secondary antibody.