XBP-1 Antibody

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ProSci
Product Code: PSI-3687
Product Group: Primary Antibodies
Supplier: ProSci
CodeSizePrice
PSI-3687-0.02mg0.02mg£150.00
Quantity:
PSI-3687-0.1mg0.1mg£449.00
Quantity:
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Overview

Host Type: Rabbit
Antibody Isotype: IgG
Antibody Clonality: Polyclonal
Regulatory Status: RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Immunohistochemistry (IHC)
  • Western Blot (WB)

Images

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<strong>Figure 1 Western Blot Validation in Human HepG2 Cell Lysate</strong><br> Loading: 15 μg of lysates per lane. Antibodies: XBP-1 3687 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. (A) the absence and (B) the presence of blocking peptide
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<strong>Figure 2 Western Blot Validation with Recombinant Protein</strong><br> Loading: 100 ng of human XBP-1 recombinant protein per lane. Antibodies: XBP-1 3687 (A: 0.5 μg/mL, B: 1 μg/mL and C: 2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Observed at around 23kD.
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<strong>Figure 3 Western Blot Validation of XBP-1 in A549 Cells</strong><br> Loading: 15 μg of A549 cell lysate Antibodies: XBP-1 3687, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane1: 0.25 μg/mL Lane2: 0.5 μg/mL Lane3: 1 μg/mL
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<strong>Figure 4 Western Blot Validation of XBP-1 in HepG2 Cells</strong><br> Loading: 15 μg of HepG2 cell lysate Antibodies: XBP-1 3687, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane1: 1 μg/mL Lane2: 2 μg/mL
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<strong>Figure 5 Immunofluorescence Validation of XBP-1 in Human HepG2 Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed HepG2 cells labeling XBP-1 with 3687 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
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<strong>Figure 6 Immunofluorescence Validation of XBP-1 in Human Pancreas Tissue</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed human pancreas tissue labeling XBP-1 with 3687 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
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<strong>Figure 7 Immunofluorescence Validation of XBP-1 in Human Liver Tissue</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed Human Liver Tissue labeling XBP-1 with 3687 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
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<strong>Figure 8 Immunofluorescence Validation of XBP-1 in Mouse Pancreas Tissue</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse pancreas tissue labeling XBP-1 with 3687 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
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<strong>Figure 9 Immunohistochemistry Validation of XBP-1 in Mouse Spleen Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-XBP-1 antibody (3687) at 2 μg/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.
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<strong>Figure 10 Immunohistochemistry Validation of XBP-1 in Mouse Testis Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-XBP-1 antibody (3687) at 2 μg/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.
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<strong>Figure 11 Immunohistochemistry Validation of XBP-1 in Rat Liver Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded Rat Liver Tissue using anti-XBP-1 antibody (3687) at 5 μg/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.
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<strong>Figure 1 Western Blot Validation in Human HepG2 Cell Lysate</strong><br> Loading: 15 μg of lysates per lane. Antibodies: XBP-1 3687 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. (A) the absence and (B) the presence of blocking peptide
<strong>Figure 2 Western Blot Validation with Recombinant Protein</strong><br> Loading: 100 ng of human XBP-1 recombinant protein per lane. Antibodies: XBP-1 3687 (A: 0.5 μg/mL, B: 1 μg/mL and C: 2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Observed at around 23kD.
<strong>Figure 3 Western Blot Validation of XBP-1 in A549 Cells</strong><br> Loading: 15 μg of A549 cell lysate Antibodies: XBP-1 3687, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane1: 0.25 μg/mL Lane2: 0.5 μg/mL Lane3: 1 μg/mL
<strong>Figure 4 Western Blot Validation of XBP-1 in HepG2 Cells</strong><br> Loading: 15 μg of HepG2 cell lysate Antibodies: XBP-1 3687, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane1: 1 μg/mL Lane2: 2 μg/mL
<strong>Figure 5 Immunofluorescence Validation of XBP-1 in Human HepG2 Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed HepG2 cells labeling XBP-1 with 3687 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
<strong>Figure 6 Immunofluorescence Validation of XBP-1 in Human Pancreas Tissue</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed human pancreas tissue labeling XBP-1 with 3687 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
<strong>Figure 7 Immunofluorescence Validation of XBP-1 in Human Liver Tissue</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed Human Liver Tissue labeling XBP-1 with 3687 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
<strong>Figure 8 Immunofluorescence Validation of XBP-1 in Mouse Pancreas Tissue</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse pancreas tissue labeling XBP-1 with 3687 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
<strong>Figure 9 Immunohistochemistry Validation of XBP-1 in Mouse Spleen Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-XBP-1 antibody (3687) at 2 μg/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.
<strong>Figure 10 Immunohistochemistry Validation of XBP-1 in Mouse Testis Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-XBP-1 antibody (3687) at 2 μg/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.
<strong>Figure 11 Immunohistochemistry Validation of XBP-1 in Rat Liver Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded Rat Liver Tissue using anti-XBP-1 antibody (3687) at 5 μg/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.

Documents

Further Information

Additional Names:
XBP-1 Antibody: XBP2, TREB5, XBP-1, XBP2, X-box-binding protein 1, Tax-responsive element-binding protein 5
Application Note:
WB: 0.25-2 μg/mL; IF: 20 μg/mL; ICC: 10 μg/mL; IHC: 2-5 μg/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.
Background:
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.
Background References:
  • Yoshida et al. Cell 2001; 107:881-91.
  • Calfon et al. Nature 2002; 415:92-6.
  • Haze et al. Mol. Cell. Biol. 1999; 10:3787-99.
  • Little et al. Crit. Rev. Eukaryot. Gene Expr. 1994; 4:1-18.
Buffer:
XBP-1 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Conjugate:
Unconjugated
DISCLAIMER:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.
Homology:
Predicted species reactivity based on immunogen sequence: Bovine: (100%)
Immunogen:
Anti-XBP-1 antibody (3687) was raised against a peptide corresponding to 17 amino acids near the amino terminus of human XBP-1.

The immunogen is located within amino acids 40 - 90 of XBP-1.
ISOFORMS:
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.
NCBI Gene ID #:
7494
NCBI Official Name:
X-box binding protein 1
NCBI Official Symbol:
XBP1
NCBI Organism:
Homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
Predicted: 28/40kD

Observed: 28/40 kD
Protein Accession #:
P17861
Protein GI Number:
60416406
Purification:
XBP-1 Antibody is affinity chromatography purified via peptide column.
Research Area:
Signal Transduction,Chemokines & Cytokines
Swissprot #:
P17861
User NOte:
Optimal dilutions for each application to be determined by the researcher.
VALIDATION:

Recombinant Protein Test (Figure 3): Anti-XBP-1 antibodies (3687) detected human XBP-1 recombinant protein at different concentrations.

Regulated expression validation (Figure 12): XBP-1 expression detected by anit-XBP-1 antibodies was up-regulated by adenovirus encoding beta-gal but was down-regulated by adenovirus encoding human GRP78 in ob/ob mice.

References

  1. Kammoun et al. GRP78 expression inhibits insulin and ER stress-induced SREBP-1c activation and reduces hepatic steatosis in mice. J Clin Invest. 2009;119(5):1201-15. PMID: PMID:19363290
  2. 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

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