Revision 7
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, IP, IHC-Bond, IHC-P

REACTIVITY:

H M Mk

SENSITIVITY:

Endogenous

MW (kDa):

90, 120

Source/Isotype:

Rabbit IgG

UniProt ID:

#P28908

Entrez-Gene Id:

943

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
IHC Leica Bond 1:100 - 1:400
Immunohistochemistry (Paraffin) 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.

For a carrier-free (BSA and azide free) version of this product see product #47448.

Specificity / Sensitivity

TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb recognizes endogenous levels of total TNFRSF8/CD30 protein. This antibody is predicted to detect CD30V, which is a truncated isoform that consists of the CD30 cytoplasmic domain. Non-specific staining was observed in the epithelium of mouse prostate tissue.

Species Reactivity:

Human, Mouse, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu500 of human TNFRSF8/CD30 protein.

Background

TNFRSF8/CD30 is a type-I transmembrane glycoprotein that is a member of the TNFR superfamily. CD30 is synthesized as a precursor protein that undergoes extensive post-translational modification before becoming embedded in the plasma membrane as a 120-kDa transmembrane protein (1,2). The expression of CD30 is upregulated in activated T cells and may trigger costimulatory signaling pathways upon its engagement (3,4). While its expression is normally restricted to subsets of activated T cells and B cells, CD30 expression is robustly upregulated in hematologic malignancies, such as Hodgkin lymphoma (HL), anaplastic large cell lymphoma (ALCL), and adult T-cell leukemia, thus making it an attractive target for therapeutic intervention (5,6). Research studies have suggested that in certain disease contexts, CD30 recruits TRAF2 and TRAF5 adaptor proteins to drive NF-kappa B activation, aberrant cell growth, and cytokine production (7-9). CD30 signaling is also regulated by TACE-dependent proteolytic cleavage of its ectodomain, which results in reduced CD30L-dependent activation of CD30+ cells (10,11).

  1. Froese, P. et al. (1987) J Immunol 139, 2081-7.
  2. Nawrocki, J.F. et al. (1988) J Immunol 141, 672-80.
  3. Del Prete, G. et al. (1995) J Exp Med 182, 1655-61.
  4. Gilfillan, M.C. et al. (1998) J Immunol 160, 2180-7.
  5. Stein, H. et al. (1985) Blood 66, 848-58.
  6. Chiarle, R. et al. (1999) Clin Immunol 90, 157-64.
  7. Horie, R. et al. (2002) Am J Pathol 160, 1647-54.
  8. Horie, R. et al. (2002) Oncogene 21, 2493-503.
  9. Horie, R. et al. (2004) Cancer Cell 5, 353-64.
  10. Hansen, H.P. et al. (2000) J Immunol 165, 6703-9.
  11. Gruss, H.J. et al. (1997) Immunol Today 18, 156-63.

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin)

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.
KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge.
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 7
#54535

TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb

Western Blotting Image 1: TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, TNFRSF8/CD30 protein is not expressed in either U-937 or HeLa cells. KARPAS-299 cell line source: Dr. Abraham Karpas at the University of Cambridge.
Western Blotting Image 2: TNFRSF8/CD30 (E4L4I) 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 TNFRSF8/CD30 protein (hCD30-Myc/DDK; +) or a construct expressing Myc/DDK-tagged full-length mouse TNFRSF8/CD30 protein (mCD30-Myc/DDK; +), using TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb (upper), DYKDDDDK Tag Antibody #2368 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 3: TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from mouse primary CD8+ T cells, EL4 cells, and NIH/3T3 cells using TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, TNFRSF8/CD30 protein is not detected in NIH/3T3 cells. CD8+ T cells were purified from mouse spleen and stimulated for 7 days using beads coated with CD3 and CD28 antibodies in the presence of mouse interleukin-2 (20 ng/mL).
Immunoprecipitation Image 1: TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb Expand Image
Immunoprecipitation of TNFRSF8/CD30 from KARPAS-299 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb. Western blot analysis was performed using TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
Immunohistochemistry Image 1: TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human Hodgkin lymphoma using TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb performed on the Leica BOND Rx. Note prominent staining of Reed-Sternberg cells.
Immunohistochemistry Image 1: TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human Hodgkin lymphoma using TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb. Note prominent staining of Reed-Sternberg cells.
Immunohistochemistry Image 2: TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human tonsil using TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb.
Immunohistochemistry Image 3: TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin lymphoma using TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb.
Immunohistochemistry Image 4: TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb.
Immunohistochemistry Image 5: TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded rhesus monkey spleen using TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb.
Immunohistochemistry Image 6: TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse spleen using TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb.
Immunohistochemistry Image 7: TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded KARPAS-299 cell pellet (left, positive) or HeLa cell pellet (right, negative) using TNFRSF8/CD30 (E4L4I) XP® Rabbit mAb.