PA5-43332

antibody from Invitrogen Antibodies

Targeting: RAB5IF C20orf24, PNAS-11, RCAF1, RIP5

Provider product page for PA5-43332

Western blot

Other assay

Antibody data

Antibody Data
Antigen structure
References [2]
Comments [0]
Validations
Western blot [1]
Other assay [2]

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Product number: PA5-43332 - Provider product page
Provider: Invitrogen Antibodies
Product name: C20orf24 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: Peptide sequence: MSGGRRKEEP PQPQLANGAL KVSVWSKVLR SDAAWEDKDE FLDVIYWFRQ
Concentration: 1 mg/mL

RAB5IF protein structure - PA5-43332 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 6 SYNJ2BP binds to specific mitochondrial mRNAs at the OMM and promotes their translation during recovery from stress. a Validation of five hits from Fig. 5d as OMM-localized RBPs by UV crosslinking-based APEX-PS. Non-RBP ACTB was used as a negative control. b Validation of RNA-binding activity using metabolic RNA labeling by 5-EU and UV crosslinking. c Confocal imaging of mitochondrial RBP orphan TAX1BP1. Anti-TOMM20 stains mitochondria and DAPI stains nuclei. Scale bars, 10 mum. d Comparison of RNA-binding activities for OMM-localized and nuclear-localized EXD2 using APEX-PS-OMM and APEX-PS-NLS. Direct streptavidin enrichment (APEX only) was performed to quantify total EXD2 protein levels in each compartment. PS only condition shows RNA-binding activity of total EXD2. e Validation of mitochondria-related SYNJ2BP clients in the absence or presence of protein translation inhibitor PUR by CLIP and qRT-PCR. IARS2 is an OMM-localized mRNA not identified to bind with SYNJ2BP. Other SYNJ2BP clients and negative controls are shown in Supplementary Fig. 10a . f Evaluation of OMM localization of SYNJ2BP clients in SYNJ2BP knockout cells by APEX-mediated proximity labeling of RNA. The OMM localization was determined by comparing APEX2-OMM with APEX2-NES labeling ( y -axis). Other SYNJ2BP clients and negative controls are shown in Supplementary Fig. 10d . g Impact of SYNJ2BP knockout on protein synthesis of SYNJ2BP-regulated clients under different cellular states. The newly synth

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 7 SYNJ2BP improves mitochondrial OXPHOS recovery and cell viability following stress. a , b Evaluation of Complex III ( a ) and IV ( b ) activities in SYNJ2BP knockout cells. c Evaluation of complex I-V assembly 12 h following treatment of HEK cells with protein translation inhibitor PUR. The quantification of each OXPHOS complex is shown below; data from three independent biological replicates. d Evaluation of cell viability 12 h following treatment of HEK cells with PUR. e , f Evaluation of protein synthesis for SYNJ2BP clients in SYNJ2BP knockout cells following heat ( e ) and sodium arsenite ( f ) stresses. The quantification of western blot data from three biological replicates is shown in Supplementary Fig. 12b-c . g Evaluation of Complex III activity in SYNJ2BP knockout cells following heat stress. h Working model of SYNJ2BP's role in retaining important (blue) mRNAs at the OMM during stress, to facilitate their rapid local translation for restoration of mitochondrial function during stress recovery (right). Two-sided Student's t -test was performed and values represent means +- SD from three biological replicates.