NFIB Polyclonal antibody

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• KD/KO Validated

NFIB Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

Cat no : 29898-1-AP

Synonyms

NF1-B, NF1 B, NF I/B, HMGIC/NFIB, CTF

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Validation Data Gallery

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  WB analysis using 29898-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 29898-1-AP (NFIB antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  IHC staining of human skin cancer using 29898-1-AP

  Immunohistochemical analysis of paraffin-embedded skin cancer slide using 29898-1-AP (NFIB antibody) at dilution of 1:200 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 29898-1-AP

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using NFIB antibody (29898-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

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  IF Staining of HepG2 using 29898-1-AP

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using NFIB antibody (29898-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

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  FC experiment of HeLa using 29898-1-AP

  1X10^6 HeLa cells were intracellularly stained with 0.4 ug Anti-Human NFIB (29898-1-AP) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Isotype Control. Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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Tested Applications

• Positive WB detected in: HUVEC cells, HeLa cells, HepG2 cells, NIH/3T3 cells
• Positive IHC detected in: human skin cancer 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: HepG2 cells, HeLa cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

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

29898-1-AP targets NFIB in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: NFIB fusion protein Ag31946
• Full Name: nuclear factor I/B
• Calculated Molecular Weight: 47 kDa
• Observed Molecular Weight: 45-65 kDa
• GenBank Accession Number: BC001283
• Gene Symbol: NFIB
• Gene ID (NCBI): 4781
• 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

NFIB, also named as Nuclear factor 1, is a 420 amino acid protein, which belongs to the CTF/NF-I family and may bind DNA as a homodimer. NFIB recognizes and binds the palindromic sequence 5'-TTGGCNNNNNGCCAA-3' present in viral and cellular promoters and in the origin of replication of adenovirus type 2. These proteins are individually capable of activating transcription and replication. NFIB also exists in multiple isoforms in prostate cancer cells, with average molecular weights of 62 kDa, 57 kDa, and 49 kDa (PMID: 32692871).