NELFB Polyclonal antibody

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• KD/KO Validated

NELFB Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, IP, ELISA

Conjugate

Unconjugated

Publications

6

Cat no : 16418-1-AP

Synonyms

NELF-B, NELF B, Negative elongation factor B, KIAA1182, Cofactor of BRCA1

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Validation Data Gallery

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  WB analysis using 16418-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 16418-1-AP (NELFB antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  WB analysis of HeLa using 16418-1-AP

  WB result of NELFB antibody (16418-1-AP; 1:10000; incubated at room temperature for 1.5 hours) with sh-Control and sh-NELFB transfected HeLa cells.

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  WB analysis of BxPC-3 using 16418-1-AP

  BxPC-3 cells were subjected to SDS PAGE followed by western blot with 16418-1-AP (NELFB antibody at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of HepG2 using 16418-1-AP

  HepG2 cells were subjected to SDS PAGE followed by western blot with 16418-1-AP (NELFB antibody at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of BxPC-3 using 16418-1-AP

  BxPC-3 cells were subjected to SDS PAGE followed by western blot with 16418-1-AP (NELFB antibody at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  IP experiment of mouse brain using 16418-1-AP

  IP result of anti-NELFB (IP:16418-1-AP, 3ug; Detection:16418-1-AP 1:500) with mouse brain tissue lysate 6000ug.

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  IHC staining of human urothelial carcinoma using 16418-1-AP

  Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma tissue slide using 16418-1-AP (NELFB 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 urothelial carcinoma using 16418-1-AP

  Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma tissue slide using 16418-1-AP (NELFB antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of BxPC-3 using 16418-1-AP

  Immunofluorescent analysis of BxPC-3 cells using 16418-1-AP (NELFB antibody) at dilution of 1:25 and Alexa Fluor 488-conjugated Goat Anti-Rabbit IgG(H+L).

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Tested Applications

• Positive WB detected in: BxPC-3 cells, HeLa cells, HepG2 cells, MCF-7 cells, rat brain tissue
• Positive IP detected in: mouse brain tissue
• Positive IHC detected in: human urothelial carcinoma 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/ICC detected in: BxPC-3 cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:6000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:200-1:1000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:10-1:100

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

• WB: See 3 publications below
• IHC: See 2 publications below
• IF: See 1 publications below

Product Information

16418-1-AP targets NELFB in WB, IHC, IF/ICC, IP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: NELFB fusion protein Ag9599
• Full Name: cofactor of BRCA1
• Calculated Molecular Weight: 580aa,66 kDa; 475aa,54 kDa
• Observed Molecular Weight: 66 kDa
• GenBank Accession Number: BC011892
• Gene Symbol: NELFB
• Gene ID (NCBI): 25920
• RRID: AB_2276536
• 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

Cofactor of BRCA1 (COBRA1) was first identified as a BRCA1-interacting protein and subsequently found to be the B subunit of the negative elongation factor complex (NELF-B). Cobra1 is essential for early embryogenesis (PMID: 19340312). The MW of this protein is 66 kDa, and this antibody specially recognises the 66 kDa protein.

Publications

Species	Application	Title
mouse	IF	Nature; The landscape of RNA Pol II binding reveals a stepwise transition during ZGA.; Authors - Bofeng Liu
mouse	WB	Genes Dev; High constitutive activity of a broad panel of housekeeping and tissue-specific cis-regulatory elements depends on a subset of ETS proteins.; Authors - Alessia Curina
human	WB	Nat Commun; ERK-mediated NELF-A phosphorylation promotes transcription elongation of immediate-early genes by releasing promoter-proximal pausing of RNA polymerase II; Authors - Seina Ohe
mouse	IHC	bioRxiv; RNA polymerase II pausing is essential during spermatogenesis for appropriate gene expression and completion of meiosis; Authors - Emily G Kaye
human	WB	J Biol Chem; An Essential Signaling Function of Cytoplasmic NELFB is Independent of RNA Polymerase II Pausing; Authors - Haihui Pan
mouse	IHC	Nat Commun; RNA polymerase II pausing is essential during spermatogenesis for appropriate gene expression and completion of meiosis; Authors - Emily G Kaye