Revision 7

#82504Store 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, W-S, IHC-P, IF-IC, FC-FP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

90

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q00987

Entrez-Gene Id:

4193

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunohistochemistry (Paraffin) 1:50 - 1:200
Immunofluorescence (Immunocytochemistry) 1:400 - 1:800
Flow Cytometry (Fixed/Permeabilized) 1:400 - 1:1600

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 #62464.

Specificity / Sensitivity

MDM2 (F7W7X) Rabbit mAb recognizes endogenous levels of total MDM2 protein. Based on the sequence of the protein antigen, this antibody is expected to detect full-length MDM2, as well as isoforms A, C, alpha, F, G, and 11. This antibody may detect isoforms A1, B, and D. By western blot, this antibody detects a band of unknown origin at approximately 60 kDa.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human MDM2 protein.

Background

MDM2, a ubiquitin ligase for p53, plays a central role in regulation of the stability of p53 (1). Akt-mediated phosphorylation of MDM2 at Ser166 and Ser186 increases its interaction with p300, allowing MDM2-mediated ubiquitination and degradation of p53 (2-4). Phosphorylation of MDM2 also blocks its binding to p19ARF, increasing the degradation of p53 (3).

  1. Haupt, Y. et al. (1997) Nature 387, 296-299.
  2. Mayo, L.D. and Donner, D.B. (2001) Proc. Natl. Acad. Sci. USA 98, 11598-11603.
  3. Zhou, B. P. et al. (2001) Nat. Cell Biol. 3, 973-981.
  4. Grossman, S. R. et al. (1998) Mol. Cell 2, 405-415.

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 W-S: Simple Western™ IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) 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.
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
#82504

MDM2 (F7W7X) Rabbit mAb

Western Blotting Image 1: MDM2 (F7W7X) Rabbit mAb Expand Image
Western blot analysis of extracts from SJSA-1, U-2 OS, and Saos-2 cells, untreated (-) or treated with Nutlin 3a (10 μM, 24 hr; +), using MDM2 (F7W7X) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). MDM2 is induced by Nutlin 3a treatment as expected. Negative expression of MDM2 protein in Saos-2 cells is consistent with the predicted expression pattern.
Western Blotting Image 1: MDM2 (F7W7X) Rabbit mAb Expand Image
Simple Western analysis of lysates (0.1 mg/mL) from SJSA cells treated with Nutlin 3a (10uM, 24hr) using MDM2 (F7W7X) Rabbit mAb #82504. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess ​​​​​​​Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Immunohistochemistry Image 1: MDM2 (F7W7X) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using MDM2 (F7W7X) Rabbit mAb.
Immunohistochemistry Image 2: MDM2 (F7W7X) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using MDM2 (F7W7X) Rabbit mAb.
Immunohistochemistry Image 3: MDM2 (F7W7X) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using MDM2 (F7W7X) Rabbit mAb.
Immunohistochemistry Image 4: MDM2 (F7W7X) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human gastric adenocarcinoma using MDM2 (F7W7X) Rabbit mAb.
Immunohistochemistry Image 5: MDM2 (F7W7X) Rabbit mAb Expand Image

Immunohistochemical analysis of paraffin-embedded normal human esophagus using MDM2 (F7W7X) Rabbit mAb.

Immunohistochemistry Image 6: MDM2 (F7W7X) Rabbit mAb Expand Image

Immunohistochemical analysis of paraffin-embedded normal human colon using MDM2 (F7W7X) Rabbit mAb.

Immunohistochemistry Image 7: MDM2 (F7W7X) Rabbit mAb Expand Image

Immunohistochemical analysis of paraffin-embedded normal human small intestine using MDM2 (F7W7X) Rabbit mAb.

Immunohistochemistry Image 8: MDM2 (F7W7X) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using MDM2 (F7W7X) Rabbit mAb (left) or an MDM2 Mouse mAb (right). These two antibodies detect unique, non-overlapping epitopes on human MDM2. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
Immunohistochemistry Image 9: MDM2 (F7W7X) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human squamous cell carcinoma of the skin using MDM2 (F7W7X) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right). 
Immunohistochemistry Image 10: MDM2 (F7W7X) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded U-2 OS cell pellets, untreated (left) or treated with Nutlin 3a (10 μM, 24 hr; right), using MDM2 (F7W7X) Rabbit mAb.
Immunofluorescence Image 1: MDM2 (F7W7X) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of U-2 OS cells, untreated (left) or treated with Nutlin 3a (10 μM, 24 hr; right), using MDM2 (F7W7X) Rabbit mAb (green), DyLight 554 Phalloidin #13054 (red), and DAPI #4083 (blue).
Flow Cytometry Image 1: MDM2 (F7W7X) Rabbit mAb Expand Image
Flow cytometric analysis of untreated U-2 OS cells (blue) and U-2 OS cells treated with Nutlin-3A (10 µM, 24 hr; green) using MDM2 (F7W7X) Rabbit mAb #82504 (solid lines) or 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 as a secondary antibody.