D4-GDI/RhoGDI2 Antibody (97A1015) - (cleavage specific)

Western Blot: D4-GDI/RhoGDI2 Antibody (97A1015) - (cleavage specific) [NB100-56725] - WB analysis: HL60 cells were treated for different time periods with 10 ng/ml of anti-Fas mAb (NB100-56725, Clone DX2). Harvested cells were lysed and resolved by SDS-PAGE and transferred onto membrane. D4-GDI/RhoGDI2 cleavage was detected.

Immunohistochemistry-Paraffin: D4-GDI/RhoGDI2 Antibody (97A1015) - (cleavage specific) [NB100-56725] - analysis of D4GD1-cv in formalin-fixed, paraffin-embedded human breast tumor tissue using an isotype control (top) and this antibody.

Flow (Intracellular): D4-GDI/RhoGDI2 Antibody (97A1015) - (cleavage specific) [NB100-56725] - Analysis using the FITC conjugate of NB100-56725. Staining of cleaved D4-GDI in untreated (left) and staurosporine-treated (right) Ramos cells using 0.5 ug of was used for this test.

Western Blot: D4-GDI/RhoGDI2 Antibody (97A1015) - (cleavage specific) [NB100-56725] - Western blot analysis of D4GDI in Jurkat cells using NB100-56556 (IMG-396A) at 2 ug/ml and NB100-56725 (IMG-160A) at 0.1 ug/ml. Cells were treated with 2 uM staurosporine for different time periods. NB100-56556 (IMG-396A) detects both the full-length and cleaved forms of D4GDI. While (NB100-56725) IMG-160A specifically detects only the cleaved form of the protein.

Flow (Intracellular): D4-GDI/RhoGDI2 Antibody (97A1015) - (cleavage specific) [NB100-56725] - HL60 cells were left untreated (green) or treated with camptothecin to induce apoptosis (red). Cells were analyzed by intracellular flow cytometry using either the (0.5 ug antibody/test/1 million cells) and ((as seen in B). this is because (cells not undergoing apoptosis) and cleaved (apoptotic cells) forms of D4-GDI.

Flow (Intracellular): D4-GDI/RhoGDI2 Antibody (97A1015) - (cleavage specific) [NB100-56725] - Analysis using the Alexa Fluor (R) 647 conjugate of NB100-56725. Staining of cleaved D4-GDI in 10^6 untreated (left) and 3 hour staurosporine-treated (right) Jurkat cells using 0.5 ug of this antibody was used for this test. Green histogram represents isotype control; red represents anti-D4-GDI antibody.

Flow (Intracellular): D4-GDI/RhoGDI2 Antibody (97A1015) - (cleavage specific) [NB100-56725] - Analysis using the FITC conjugate of NB100-56725. Staining of cleaved D4-GDI in 10^6 camptothecin-treated (red) and untreated (green) HL60 cells using 0.5 ug of NB100-56060. The shaded histogram represents cells without antibody.

• Reactivity: Hu, Mu
• Applications: WB, Flow, Flow-IC, IHC, IHC-P, IP
• Clone: 97A1015
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 0.5 mg/ml

D4-GDI/RhoGDI2 Antibody (97A1015) - (cleavage specific) Summary

• Immunogen: A synthetic peptide corresponding to the region of the Fas-induced cleavage site of human D4-GDI/RhoGDI2 was used as the immunogen.
• Specificity: In anti-Fas treated Jurkat, an approx. 23 kDa band should be observed.
• Isotype: IgG2b Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: ARHGDIB
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Western Blot 2 ug/ml
  • Flow Cytometry 0.5-1 ug/10^6 cells
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Paraffin 1:10-1:500
  • Immunoprecipitation 1-2 ug/ml
  • Flow (Intracellular) 0.5-1 ug/10^6 cells
• Control:

  D4-GDI/RhoGDI2 Lysate (NBL1-07674)

Reactivity Notes

Cross reacts with Human and Mouse.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS containing 0.05% BSA
• Preservative: 0.05% Sodium Azide
• Concentration: 0.5 mg/ml
• Purity: Protein G purified

Alternate Names for D4-GDI/RhoGDI2 Antibody (97A1015) - (cleavage specific)

• ARHGDIB Rho GDP dissociation inhibitor (GDI) beta
• D4
• GDIA2
• GDID4
• LYGDI
• Ly-GDI
• RAP1GN1
• RhoGDI2

Background

D4-GDI (GDP dissociation inhibitor) is a negative regulator of the ras-related Rho family of GTPases. Since the Rho GTPases promote cytoskeletal and membrane changes associated with apoptotic cell death, the removal of the D4-GDI block through its cleavage is important for inducing apoptosis. Caspase-3 cleaves the 28 kD mature form of D4-GDI to give a 23 kD and 5 kD size fragment. The 23 kD fragment then translocates to the nucleus. The mechanisms involving cleavage of D4-GDI with apoptosis are not presently known. Activation of the Jun N-terminal kinase, a regulator of apoptosis, may be one of the mechanisms.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for D4-GDI/RhoGDI2 (cleavage specific) Antibody (NB100-56725)(16)

Publications using NB100-56725

• Nishtala K, Phong TQ, Steil L et al. Proteomic analyses of age related changes in A.BY/SnJ mouse hearts. Proteome Sci Proteome Sci [PMID: 23816347] (IHC-P, Mouse)
• Okamoto M, Koga S, Tatsuka M. Differential regulation of caspase-9 by ionizing radiation- and UV-induced apoptotic pathways in thymic cells. Mutat Res. 2010 Jun 1 [PMID: 20346366] (WB, Mouse)
  
  Details:
  D4-GDI (IMG-160A). WB: Mouse lymphoma 3SB cell line subject to either ionizing radiation or UV light, Fig 2D.
• Hu LD, Zou HF, Zhan SX, Cao KM. Biphasic expression of RhoGDI2 in the progression of breast cancer and its negative relation with lymph node metastasis. Oncol Rep. 2007 Jun [PMID: 17487395] (WB)
  
  Details:
  IHC (P): Fig 1 and Table I (human breast tumor and normal adjacent tissues (Fig 1 and Table 1); WB: Fig 2A (human breast cancer cell lines).
• Jenkins CE, Swiatoniowski A, Power MR, Lin TJ. Pseudomonas aeruginosa-induced human mast cell apoptosis is associated with up-regulation of endogenous Bcl-xS and down-regulation of Bcl-xL. J Immunol. 2006 Dec 1 [PMID: 17114473]
• Wang Z, Zhang X, Shen P et al. A variant of estrogen receptor-{alpha}, hER-{alpha}36: transduction of estrogen- and antiestrogen-dependent membrane-initiated mitogenic signaling. Proc Natl Acad Sci U S A. 2006 Jun 13 [PMID: 16754886] (WB)
  
  Details:
  WB: HEk293 whole cell lysates (W), plasma membrane (M), cytosolic (C), and nuclear (N) fractions, Fig 3.
• Jenkins CE, Swiatoniowski A, Issekutz AC, Lin TJ. Pseudomonas aeruginosa exotoxin A induces human mast cell apoptosis by a caspase-8 and -3-dependent mechanism. J Biol Chem. 2004 Aug 27 [PMID: 15205454]
• Liu CY, Lee CF, Hong CH, Wei YH. Mitochondrial DNA mutation and depletion increase the susceptibility of human cells to apoptosis. Ann N Y Acad Sci. 2004 Apr [PMID: 15126291]
• Ko CH, Shen SC, Chen YC. Hydroxylation at C4' or C6 is essential for apoptosis-inducing activity of flavanone through activation of the caspase-3 cascade and production of reactive oxygen species. Free Radic Biol Med. 2004 Apr 1 [PMID: 15019974]
• Chen YC, Shen SC, Lin HY. Rutinoside at C7 attenuates the apoptosis-inducing activity of flavonoids. Biochem Pharmacol. 2003 Oct 1 [PMID: 14505793]
• Shen SC, Chen YC, Hsu FL, Lee WR. Differential apoptosis-inducing effect of quercetin and its glycosides in human promyeloleukemic HL-60 cells by alternative activation of the caspase 3 cascade. J Cell Biochem. 2003 Aug 1 [PMID: 12874837]
• Chen YC, Shen SC, Lee WR et al. Emodin induces apoptosis in human promyeloleukemic HL-60 cells accompanied by activation of caspase 3 cascade but independent of reactive oxygen species production. Biochem Pharmacol. 2002 Dec 15 [PMID: 12445860]
• Gildea JJ, Seraj MJ, Oxford G et al. RhoGDI2 is an invasion and metastasis suppressor gene in human cancer. Cancer Res. 2002 Nov 15 [PMID: 12438227]
• Sato S, Fujita N, Tsuruo T. Interference with PDK1-Akt survival signaling pathway by UCN-01 (7-hydroxystaurosporine). Oncogene. 2002 Mar 7 [PMID: 11896604]
• Yamaguchi H, Paranawithana SR, Lee MW et al. Epothilone B analogue (BMS-247550)-mediated cytotoxicity through induction of Bax conformational change in human breast cancer cells. Cancer Res. 2002 Jan 15 [PMID: 11809697]
• Yamaguchi H, Wang HG. The protein kinase PKB/Akt regulates cell survival and apoptosis by inhibiting Bax conformational change. Oncogene. 2001 Nov 22 [PMID: 11753656]
• Cuddeback SM, Yamaguchi H, Komatsu K et al. Molecular cloning and characterization of Bif-1. A novel Src homology 3 domain-containing protein that associates with Bax. J Biol Chem. 2001 Jun 8 [PMID: 11259440]

Review for D4-GDI/RhoGDI2 (cleavage specific) Antibody (NB100-56725) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Anonymous WB Human 07/18/2018

Summary

• Application: Western Blot
• Sample Tested: HT-29 human colon adenocarcinoma cell line
• Species: Human
• Lot: AB021110A-04

Comments

• Comments: HT-29 cells were treated with etoposide (100 ug/ml) for indicated time points. Total cell lysates were subjected to western blot. PVDF membrane were probed with 2 um/ml D4-GDI/RhoGDI2 (cleavage specific) Antibody (NB100-56725). A specific band was detected for cleaved RhoGDI2 at approximately 23 kDa. This experiment was conducted under reducing conditions.

Bioinformatics

• Gene Symbol: ARHGDIB