XBP-1 Antibody

blue ice or RT

Predicted species reactivity based on immunogen sequence: Bovine: (100%)

WB: 0.25-2 μ g/mL; IF: 20 μ g/mL; ICC: 10 μ g/mL; IHC: 2-5 μ g/mL.

Antibody validated: Western Blot in human samples; Immunofluorescence in human and rat samples; Immunocytochemistry in human samples; Immunohistochemistry in human, mouse, and rat samples. All other applications and species not yet tested.

Cat. No. 17-011 - Hep G2 Cell Slide

Predicted: 28/40kD

Observed: 28/40 kD

Human XBP-1 has two isoforms, including isoform 1 (261aa, 29kD) and isoform 2 (376aa, 40kD). Mouse XBP-1 also has two isoforms, including isoform 1 (267aa, 29.6kD) and isoform 2 (371aa, 40kD). And Rat XBP-1 has two isoforms as well, including isoform 1 (267aa, 30kD) and isoform 2 (371aa, 40kD). 3687 can detect human, mouse and rat.

Polyclonal

Unconjugated

1 mg/mL

Cat. No. 3687P – XBP-1 Peptide

None

P17861

7494

XBP1

Homo sapiens

XBP-1 Antibody: X box binding protein 1 (XBP-1) is a key protein in the mammalian unfolded protein response (UPR) that protects the cell against the stress of malfolded proteins in the endoplasmic reticulum (ER). Upon sensing unfolded proteins, an ER transmembrane endonuclease and kinase termed IRE1p is activated and excises an intron from XBP-1 mRNA. The spliced XBP-1 mRNA results in a 371 amino acid protein (XBP-1s) which is then translocated to the nucleus where it binds to the regulatory elements of downstream genes. Together with other UPR transcription factors such as ATF6, XBP-1 stimulates the production of ER stress proteins including the ER resident protein chaperones glucose regulated protein (GRP) 78 and GRP94.

Calfon et al. Nature 2002; 415:92-6.

Little et al. Crit. Rev. Eukaryot. Gene Expr. 1994; 4:1-18.

Lerat et al. Hepatitis C virus proteins induce lipogenesis and defective triglyceride secretion in transgenic mice. J Biol Chem. 2009; 284(48):33466-74. PMID: PMID: 19808675

Figure 2 Western Blot Validation with Recombinant Protein
Loading: 100 ng of human XBP-1 recombinant protein per lane.Antibodies: XBP-1 3687 (A: 0.5 μ g/mL, B: 1 μ g/mL and C: 2 μ g/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.Observed at around 23kD.

Figure 4 Western Blot Validation of XBP-1 in HepG2 Cells
Loading: 15 μ g of HepG2 cell lysateAntibodies: XBP-1 3687, 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.Lane1: 1 μ g/mLLane2: 2 μ g/mL

Figure 6 Immunofluorescence Validation of XBP-1 in Human Pancreas Tissue
Immunofluorescent analysis of 4% paraformaldehyde-fixed human pancreas tissue labeling XBP-1 with 3687 at 20 μ g/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

Figure 8 Immunofluorescence Validation of XBP-1 in Mouse Pancreas Tissue
Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse pancreas tissue labeling XBP-1 with 3687 at 20 μ g/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

Figure 10 Immunohistochemistry Validation of XBP-1 in Mouse Testis Tissue
Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-XBP-1 antibody (3687) at 2 μ g/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4 ˚ C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.