C9orf72 Monoclonal antibody

C9orf72 Monoclonal Antibody for WB, IHC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG2a

Reactivity

human, mouse, rat, pig

Applications

WB, IHC, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

2F9A2

Cat no : 67824-1-Ig

Synonyms

Guanine nucleotide exchange factor C9orf72, DENNL72, DENND9, C9RANT, 2F9A2

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Validation Data Gallery

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  WB analysis using 67824-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 67824-1-Ig (C9orf72 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control.

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  WB analysis of Neuro-2a using 67824-1-Ig

  Neuro-2a cells were subjected to SDS PAGE followed by western blot with 67824-1-Ig (C9orf72 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  WB analysis of PC-12 using 67824-1-Ig

  PC-12 cells were subjected to SDS PAGE followed by western blot with 67824-1-Ig (C9orf72 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of C6 using 67824-1-Ig

  C6 cells were subjected to SDS PAGE followed by western blot with 67824-1-Ig (C9orf72 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  WB analysis of pig brain using 67824-1-Ig

  pig brain tissue were subjected to SDS PAGE followed by western blot with 67824-1-Ig (C9orf72 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of rat brain using 67824-1-Ig

  rat brain tissue were subjected to SDS PAGE followed by western blot with 67824-1-Ig (C9orf72 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse brain using 67824-1-Ig

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 67824-1-Ig (C9orf72 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HEK-293 using 67824-1-Ig

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 67824-1-Ig (C9orf72 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of U-87 MG using 67824-1-Ig

  U-87 MG cells were subjected to SDS PAGE followed by western blot with 67824-1-Ig (C9orf72 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  IHC staining of mouse lung using 67824-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse lung tissue slide using 67824-1-Ig (C9orf72 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse intestine using 67824-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse intestine tissue slide using 67824-1-Ig (C9orf72 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat kidney using 67824-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat kidney tissue slide using 67824-1-Ig (C9orf72 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  FC experiment of HEK-293T using 67824-1-Ig

  1X10^6 HEK-293T cells were intracellularly stained with 0.4 ug Anti-Human C9orf72 (67824-1-Ig, Clone:2F9A2) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG2a Isotype Control (C1.18.4) (65208-1-Ig, Clone: C1.18.4) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: pig brain tissue, HEK-293 cells, U-87 MG cells, Neuro-2a cells, PC-12 cells, C6 cells, rat brain tissue, mouse brain tissue
• Positive IHC detected in: mouse lung tissue, mouse intestine, rat kidney tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive FC (Intra) detected in: HEK-293T cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:6000
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

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.

Product Information

67824-1-Ig targets C9orf72 in WB, IHC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat, pig samples.

• Tested Reactivity: human, mouse, rat, pig
• Host / Isotype: Mouse / IgG2a
• Class: Monoclonal
• Type: Antibody
• Immunogen: C9orf72 fusion protein Ag21080
• Full Name: chromosome 9 open reading frame 72
• Calculated Molecular Weight: 481 aa, 54 kDa
• Observed Molecular Weight: 50 kDa
• GenBank Accession Number: BC020851
• Gene Symbol: C9orf72
• Gene ID (NCBI): 203228
• RRID: AB_2918586
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A 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.