PA5-42646

antibody from Invitrogen Antibodies

Targeting: RHOT1 ARHT1, FLJ11040, MIRO-1, MIRO1

Provider product page for PA5-42646

Western blot

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [2]
Comments [0]
Validations
Other assay [2]

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Product number: PA5-42646 - Provider product page
Provider: Invitrogen Antibodies
Product name: RHOT1 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: Peptide sequence: MKKDVRILLV GEPRVGKTSL IMSLVSEEFP EEVPPRAEEI TIPADVTPER Sequence homology: Cow: 100%; Dog: 100%; Guinea Pig: 100%; Horse: 100%; Human: 100%; Mouse: 100%; Rabbit: 100%; Rat: 100%; Zebrafish: 93%
Reactivity: Human
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 0.5 mg/mL
Storage: -20° C, Avoid Freeze/Thaw Cycles

RHOT1 protein structure - PA5-42646 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 2 Immunoprecipitation of Miro1 with Nbs. (A) For immunoprecipitation with immobilized Nbs (nanotraps), soluble protein fraction of HEK293 cells transiently expressing GFP-Miro1 or GFP as control, was adjusted to 2 mg/ml and incubated with equal amounts of nanotraps. Input (IN, 1% of total) and bound (20% of total) fractions were subjected to SDS-PAGE followed by immunoblot analysis using antibodies specific for GFP (upper panel) and GAPDH (lower panel). As positive control GFP-nanotrap and as negative a non-specific (NC) nanotrap were used. (B) Immunoprecipitation from non-transfected HEK293 as described in (A) were performed. Input and bound fractions were analysed with an anti-Miro1 antibody. As positive control anti-Miro1 IgG immobilized on Protein A/G sepharose and as negative control a non-specific (NC) nanotrap was used. For densitometric evaluation immunoblot signals of endogenous Miro1 in the corresponding bound fractions were normalized to the Miro1 signal in the input, which was set to 1. Shown are the mean signals from three independent biological replicates +-SD. (C) Immunofluorescence (IF) detection of GFP-Miro1 in fixed and permeabilized HeLa cells after staining with Miro1-Nbs as primary labelling probes. Representative confocal laser scanning (CLSM) images are shown of each individual Nb detected with anti-VHH antibody labelled with Cy5 (middle row). As positive control, transfected cells were stained with anti-Miro1 antibody followed by detection with

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 5 Proteomic analysis of Miro1 capture. (A) For comparable immunoprecipitation soluble protein fraction of HEK293 cells were incubated either with monovalent Nbs either chemically coupled to NHS sepharose (M41 NHS , M114 NHS ) or the bivalent formats, which were site specifically conjugated to agarose particles by sortagging and click chemistry (bivM41 sort , bivM114 sort ). Input and bound fractions were analysed with an anti-Miro1 antibody. As positive control anti-Miro1 IgG immobilized on Protein A/G sepharose and as negative control a non-specific bivalent and site specifically conjugated nanotrap (bivNC sort ) was used. Shown is a representative immunoblot stained with an anti-Miro1 antibody. For densitometric evaluation immunoblot signals of endogenous Miro1 in the corresponding bound fractions were normalized to the Miro1 signal in the input, which was set to 1. Shown are the mean signals from three independent biological replicates +-SD. (B) Capture efficiency by mono- and bivalent nanotraps. Averaged iBAQ (intensity based absolute quantification) values for Miro1 isoform 2 (white circles) and isoform 3 (black circles) of three biological replicates are shown. (C) Classification of Miro1 interactor based on STRING database. Class 1: direct interactor, confidence score >0.9; Class 2: direct interactor, confidence score 0.9.