Revision 4

#98785Store 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, IP, IHC-Bond, IHC-P, IF-IC, FC-FP, FC-L

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

35-50

Source/Isotype:

Mouse IgG2a

UniProt ID:

#P43489

Entrez-Gene Id:

7293

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
IHC Leica Bond 1:400
Immunohistochemistry (Paraffin) 1:400
Immunofluorescence (Immunocytochemistry) 1:1000
Flow Cytometry (Fixed/Permeabilized) 1:200
Flow Cytometry (Live) 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 #99419.

Specificity / Sensitivity

OX40 (ACT35) Mouse mAb recognizes endogenous levels of total OX40 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with HuT 102 cell line.

Background

OX40 (TNFRSF4, CD134) is a member of the tumor necrosis factor (TNF) receptor superfamily that regulates T cell activity and immune responses. The OX40 protein contains four cysteine rich domains, a transmembrane domain, and a cytoplasmic tail containing a QEE motif (1,2). OX40 is primarily expressed on activated CD4+ and CD8+ T-cells, while the OX40 ligand (OX40L, TNFSF4, CD252) is predominantly expressed on activated antigen presenting cells (3-7). The engagement of OX40 with OX40L leads to the recruitment of TNF receptor-associated factors (TRAFs) and results in the formation of a TCR-independent signaling complex. One component of this complex, PKCθ, activates the NF-κB pathway (2,8). OX40 signaling through Akt can also enhance TCR signaling directly (9). Research studies indicate that the OX40L-OX40 pathway is associated with inflammation and autoimmune diseases (10). Additional research studies show that OX40 agonists augment anti-tumor immunity in several cancer types (11,12).

  1. Croft, M. (2010) Annu Rev Immunol 28, 57-78.
  2. So, T. and Croft, M. (2012) Front Immunol 3, 133.
  3. Paterson, D.J. et al. (1987) Mol Immunol 24, 1281-90.
  4. Mallett, S. et al. (1990) EMBO J 9, 1063-8.
  5. Dürkop, H. et al. (1995) Br J Haematol 91, 927-31.
  6. Godfrey, W.R. et al. (1994) J Exp Med 180, 757-62.
  7. Al-Shamkhani, A. et al. (1997) J Biol Chem 272, 5275-82.
  8. So, T. et al. (2011) Proc Natl Acad Sci U S A 108, 2903-8.
  9. So, T. and Croft, M. (2013) Front Immunol 4, 139.
  10. Gough, M.J. and Weinberg, A.D. (2009) Adv Exp Med Biol 647, 94-107.
  11. Weinberg, A.D. et al. (2011) Immunol Rev 244, 218-31.
  12. Linch, S.N. et al. (2015) Front Oncol 5, 34.

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) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) FC-L: Flow Cytometry (Live)

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.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
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
#98785

OX40 (ACT35) Mouse mAb

Western Blotting Image 1: OX40 (ACT35) Mouse mAb Expand Image
Western blot analysis of extracts from HuT 102, human CD4+ T, and Jurkat cells using OX40 (ACT35) Mouse mAb (upper) and β-actin (D6A8) Rabbit mAb #8457 (lower). CD4+ T cells were purified from blood, and stimulated for 9 days using beads coated with CD3 and CD28 antibodies in the presence of Interleukin-2 (hIL-2) #8907 (20 ng/ml).
Immunoprecipitation Image 1: OX40 (ACT35) Mouse mAb Expand Image
Immunoprecipitation of OX40 protein from HuT 102 cell extracts. Lane 1 is 10% input, lane 2 is Mouse (G3A1) mAb IgG1 Isotype Control #5415, and lane 3 is OX40 (ACT35) Mouse mAb. Western blot analysis was performed using

OX40 (ACT35) Mouse mAb.

Immunohistochemistry Image 1: OX40 (ACT35) Mouse mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human gastric adenocarcinoma using OX40 (ACT35) Mouse mAb performed on the Leica® BOND Rx.
Immunohistochemistry Image 2: OX40 (ACT35) Mouse mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma using OX40 (ACT35) Mouse mAb performed on the Leica® BOND Rx.
Immunohistochemistry Image 1: OX40 (ACT35) Mouse mAb Expand Image
Immunohistochemical analysis of paraffin-embedded HuT 102 cell pellet (left, positive) or Jurkat cell pellet (right, negative) using OX40 (ACT35) Mouse mAb.
Immunohistochemistry Image 2: OX40 (ACT35) Mouse mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human squamous cell carcinoma of the lung using OX40 (ACT35) Mouse mAb.
Immunohistochemistry Image 3: OX40 (ACT35) Mouse mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human tonsil using OX40 (ACT35) Mouse mAb (left) compared to concentration matched Mouse (G3A1) IgG1 Isotype Control #5415 (right).
Immunofluorescence Image 1: OX40 (ACT35) Mouse mAb Expand Image
Confocal immunofluorescent analysis of HuT 102 cells (left, positive) and Jurkat cells (right, negative) using OX40 (ACT35) Mouse mAb (green). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Flow Cytometry Image 1: OX40 (ACT35) Mouse mAb Expand Image
Flow cytometric analysis of Jurkat cells (blue) and HuT 102 cells (green) using OX40 (ACT35) Mouse mAb (solid lines) or a concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.