C9orf72 Polyclonal antibody

• Featured Product
• KD/KO Validated

C9orf72 Polyclonal Antibody for IF, IHC, WB,ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse and More (1)

Applications

WB, IHC, IF, ELISA

Conjugate

Unconjugated

Publications

6

Cat no : 25757-1-AP

Synonyms

C9orf72, C9RANT

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Validation Data Gallery

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  WB analysis of HEK-293 using 25757-1-AP

  WB result of C9orf72 antibody (25757-1-AP, 1:250 incubated at RT for 1 hour) with si-C9orf72 HEK293 cells, C9orf72 CRISPR KO HEK293 cells; normal HEK293 cells and non-targeting control (NTC) siRNA transfected HEK293 cells as control. The ~50 kDa is the C9orf72 Long isoform. (Data from Dr Chris Webster, Postdoc in Kurt de Vos’s group at SITRAN).

• 

  WB analysis of mouse brain using 25757-1-AP

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 25757-1-AP (C9orf72 Antibody) at dilution of 1:1500 incubated at room temperature for 1.5 hours.

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  WB analysis of SH-SY5Y using 25757-1-AP

  SH-SY5Y cells were subjected to SDS PAGE followed by western blot with 25757-1-AP (C9orf72 Antibody) at dilution of 1:300 incubated at 4 degree celsius over night.

• 

  IHC staining of human gliomas using 25757-1-AP

  Immunohistochemical analysis of paraffin-embedded human gliomas tissue slide using 25757-1-AP (C9orf72 antibody) at dilution of 1:1000 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human gliomas using 25757-1-AP

  Immunohistochemical analysis of paraffin-embedded human gliomas tissue slide using 25757-1-AP (C9orf72 antibody) at dilution of 1:1000 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human testis using 25757-1-AP

  Immunohistochemical analysis of paraffin-embedded human testis tissue slide using 25757-1-AP (C9orf72 Antibody) at dilution of 1:50 (under 10x lens).

• 

  IHC staining of human testis using 25757-1-AP

  Immunohistochemical analysis of paraffin-embedded human testis tissue slide using 25757-1-AP (C9orf72 Antibody) at dilution of 1:50 (under 40x lens).

• 

  IF Staining of mouse brain using 25757-1-AP

  Immunofluorescent analysis of (4% PFA) fixed mouse brain tissue using 25757-1-AP (C9orf72 antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

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Tested Applications

• Positive WB detected in: mouse brain tissue, HEK-293 cells, SH-SY5Y cells
• Positive IHC detected in: human gliomas tissue, human testis tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF detected in: mouse brain tissue

Recommended dilution

• Western Blot (WB): WB : 1:300-1:1500
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF): IF : 1:50-1:500

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 1 publications below
• WB: See 5 publications below
• IHC: See 1 publications below
• IF: See 2 publications below

Product Information

The immunogen of 25757-1-AP is C9orf72 Fusion Protein expressed in E. coli.

• Tested Reactivity: human, mouse
• Cited Reactivity: human, rat
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: C9orf72 fusion protein Ag22723
• Full Name: chromosome 9 open reading frame 72
• Calculated Molecular Weight: 481 aa, 54 kDa
• Observed Molecular Weight: 25-30 kDa
• GenBank Accession Number: BC068445
• Gene Symbol: C9orf72
• Gene ID (NCBI): 203228
• RRID: AB_2784548
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

C9ORF72 has a domain whith polymorphic hexanucleotide repeat (GGGGCC). The C9ORF72-hexanucleotide repeat expansions have been recently identified as genetic markers in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). The C9ORF72 repeat expansions may indicate a worse prognosis in ALS. Human C9ORF72 has some isoforms with MW 54-60 kDa and 25-30 kDa. Mouse C9orf72 has some isoforms with MW 50-60 kDa and 35 kDa. This antibody detects the N-terminal of C9orf72.

Publications

Species	Application	Title
human	IHC	Nat Med; Haploinsufficiency leads to neurodegeneration in C9ORF72 ALS/FTD human induced motor neurons.; Authors - Yingxiao Shi
human	WB	Acta Neuropathol; An interaction between synapsin and C9orf72 regulates excitatory synapses and is impaired in ALS/FTD.; Authors - Claudia S Bauer
human	WB,IF	Elife; Implementation of an antibody characterization procedure and application to the major ALS/FTD disease gene C9ORF72.; Authors - Carl Laflamme
human,rat	WB	Theranostics; C9orf72 regulates the unfolded protein response and stress granule formation by interacting with eIF2α; Authors - Wenzhong Zheng
human	WB	Front Cell Neurosci; Loss of TMEM106B exacerbates C9ALS/FTD DPR pathology by disrupting autophagosome maturation; Authors - Claudia S Bauer
human	WB,IF	EMBO Rep; C9orf72 protein quality control by UBR5-mediated heterotypic ubiquitin chains; Authors - Julia Jülg ; KO Validated

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