Rabex5 Antibody - BSA Free

Western Blot: Rabex5 Antibody [NBP1-49938] - Detection of Human and Mouse RABGEF1 by Western Blot (h&m) and Immunoprecipitation (h). Samples: Whole cell lysate from HeLa (5, 15 and 50 mcg for WB; 1 mg for IP, 20% of IP loaded), 293T (T; 50 mcg) and mouse NIH3T3 (M; 50 mcg) cells. Antibodies: Affinity purified rabbit anti-RABGEF1 antibody used for WB at 0.1 mcg/ml (A) and 1 mcg/ml (B) and used for IP at 3 mcg/mg lysate. RABGEF1 was also immunoprecipitated by rabbit anti-RABGEF1 antibody which recognizes a downstream epitope. For blotting immunoprecipitated RABGEF1 was used. Detection: Chemiluminescence with exposure times of 3 minutes (A) and 30 seconds (B).

Western Blot: Rabex5 Antibody [NBP1-49938] - RABGEF1 is important for mitochondrial clearance. YFP-Parkin stably expressing WT and RABGEF1-mAID HCT116 cells pre-treated with IAA were treated with valinomycin for the indicated times. Total cell lysates were analyzed by immunoblotting. Image collected and cropped by CiteAb from the following publication (//elifesciences.org/articles/31326), licensed under a CC-BY license.

Immunohistochemistry-Paraffin: Rabex5 Antibody [NBP1-49938] - Section of human lung carcinoma. Antibody: Affinity purified rabbit anti- RABGEF1 used at a dilution of 1:1,000 (1ug/ml). Detection: DAB

Western Blot: Rabex5 Antibody [NBP1-49938] - RABGEF1 recruitment to mitochondria during mitophagy.(A) The indicated cells were treated with DMSO or valinomycin for 3 hr followed by immunostaining. Bars, 10 μm. Graphs for quantification of RABGEF1 recruitment to mitochondria were shown below the images. None, partial & complete denote that GFP-mRABGEF1 signals were overlapped with no, some of, & all mitochondria, respectively. The error bars represent mean ±SE & over 100 cells were counted in each of three separate wells. (B) WT & TBC1D15/17 DKO HCT116 cells stably expressing mCherry-Parkin & GFP-mRABGEF1 were treated with DMSO or valinomycin for 3 hr. GFP-mRABGEF1 signals were enhanced by immunostaining with anti-GFP antibody. Bars, 10 μm. (C) Total cell lysates in (B) were analyzed by immunoblotting. * & # denote ubiquitinated forms & truncated forms, respectively.10.7554/eLife.31326.031Figure 8—figure supplement 1—source data 1.This excel file contains quantification of RABGEF1 (WT & Y26A/A58D mutant) recruitment to mitochondria in HCT116 (WT & TBC1D15/17 DKO) cells.This excel file contains quantification of RABGEF1 (WT & Y26A/A58D mutant) recruitment to mitochondria in HCT116 (WT & TBC1D15/17 DKO) cells. Image collected & cropped by CiteAb from the following publication (//elifesciences.org/articles/31326), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Rabex5 Antibody [NBP1-49938] - RABGEF1 is important for mitochondrial clearance.(A) WT & RABGEF1-mAID HCT116 cells were treated with or without IAA for 16 hr. Total cell lysates were analyzed by immunoblotting. (B) Quantification of Parkin recruitment to mitochondria in WT & RABGEF1-mAID HCT116 cells after 3 hr of valinomycin treatment. Partial & complete denote that YFP-Parkin signals were overlapped with some of & all mitochondria, respectively. (C) YFP-Parkin stably expressing WT & RABGEF1-mAID HCT116 cells pre-treated with IAA were treated with valinomycin for the indicated times. Total cell lysates were analyzed by immunoblotting. (D) WT & RABGEF1-mAID HCT116 cells stably expressing YFP-Parkin & mt-mKeima were treated with IAA for 16 hr followed by DMSO or OAQ for 6 hr & subjected to FACS analysis. Plots are representative of n = 3 experiments. (E) Quantification of mitophagy in (D). Error bars represent mean ±SE of three independent experiments. Statistical differences were determined by student’s t-test. *p<0.05.10.7554/eLife.31326.035Figure 9—source data 1.Quantification of YFP-Parkin recruitment to mitochondria in RABGEF1-mAID HCT116 & the corresponding WT cells during mitophagy.10.7554/eLife.31326.036Figure 9—source data 2.Quantification of mitophagy using mt-mKeima & FACS analysis.Quantification of YFP-Parkin recruitment to mitochondria in RABGEF1-mAID HCT116 & the corresponding WT cells during mitophagy.Quantification of mitophagy using mt-mKeima & FACS analysis. Image collected & cropped by CiteAb from the following publication (//elifesciences.org/articles/31326), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu, Bv
• Applications: WB, IHC, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

Rabex5 Antibody - BSA Free Summary

• Immunogen: The immunogen for this product maps to a region between residue 175 and 225 of human RAB Guanine Nucleotide Exchange Factor (GEF) 1 using the numbering given in entry BAC87138.1 (GeneID 27342).
• Predicted Species: Bovine (100%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: RABGEF1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:500- 1:2000
  • Immunoprecipitation 2-5 ug/mg lysate
  • Western Blot 1:2000-1:10000
• Application Notes: Epitope retrieval with citrate buffer pH6.0 is recommended for FFPE tissue sections.

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: Tris-Citrate/Phosphate (pH 7.0 - 8.0)
• Preservative: 0.09% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for Rabex5 Antibody - BSA Free

• FLJ32302
• RAB guanine nucleotide exchange factor (GEF) 1
• rab5 GDP/GTP exchange factor
• Rabaptin-5-associated exchange factor for Rab5
• Rabex-5
• RABEX5rabex-5
• RAP1

Background

RABGEF1 forms a complex with rabaptin-5 (RABPT5; MIM 603616) that is required for endocytic membrane fusion, and it serves as a specific guanine nucleotide exchange factor for RAB5 (RAB5A; MIM 179512) (Horiuchi et al., 1997 [PubMed 9323142]).[supplied by OMIM]

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 Rabex5 Antibody (NBP1-49938)(2)

Publications using NBP1-49938

• Yamano K, Wang C, Sarraf SA et al. Endosomal Rab cycles regulate Parkin-mediated mitophagy Elife 2018-01-23 [PMID: 29360040] (WB, Human)
• Lucitti JL, Sealock R, Buckley BK et al. Variants of Rab GTPase-Effector Binding Protein-2 Cause Variation in the Collateral Circulation and Severity of Stroke. Stroke. 2016-12-01 [PMID: 27811335]

Bioinformatics

• Gene Symbol: RABGEF1