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<strong>Figure 1 Immunofluorescence Validation of XBP-1 in HeLa Cells</strong><br>Immunofluorescent analysis of methanol-fixed HeLa cells labeling XBP-1 with 3687 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/1000 dilution (red) and DAPI staining (blue). Alpha tubulin was stained with anti-alpha tubulin antibody following by goat anti-mouse IgG secondary antibody (green). Images were captured with confocal microscopy.
<strong>Figure 2. XBP-1 KO Validation in 293T Cells</strong><br>Loading: 10 μg of lysate Antibodies: XBP-1 3687, 4 μg/mL and beta-actin 3779-1301, 1μg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 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 4 Western Blot Validation of XBP-1 in HepG2 Cells</strong><br>Loading: 15 μg 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 μg/mLLane2: 2 μg/mL
<strong>Figure 5 Western Blot Validation in Mouse Pancreas Tissue</strong><br>Loading: 15 μg of lysates per lane.Antibodies: XBP-1, 3687 (4 μg /mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 6 Western Blot Validation in Rat Liver Tissue 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.(1) the absence and (2) the presence of blocking peptide
<strong>Figure 7 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 8 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 9 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 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 Mouse Spleen Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-XBP-1 antibody (3687) at 1 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 12 Immunohistochemistry Validation of XBP-1 in Rat Spleen Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded rat spleen tissue using anti-XBP-1 antibody (3687) at 1 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.
Figure 1 Immunofluorescence Validation of XBP-1 in HeLa Cells
Immunofluorescent analysis of methanol-fixed HeLa cells labeling XBP-1 with 3687 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/1000 dilution (red) and DAPI staining (blue). Alpha tubulin was stained with anti-alpha tubulin antibody following by goat anti-mouse IgG secondary antibody (green). Images were captured with confocal microscopy.
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XBP-1 Antibody

CATALOG NUMBER: 3687

Clonality:
Polyclonal
Tested Applications:
ELISA, ICC, IF, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
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.
Conjugate:
Unconjugated
Tested Applications:
ELISA, ICC, IF, IHC-P, WB
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.
Positive Control 1:
Cat. No. 1414 – Mouse Pancreas Tissue Lysate
Positive Control 2:
Cat. No. 1464 – Rat Liver Tissue Lysate
Predicted Molecular Weight:
Predicted: 28/40kD
Observed: 28/40 kD
Validation:

KO Validation (Figure 2): XBP-1 expression detected by anti-XBP-1 antibodies was observed in wild type HeLa cells, but not in the XBP-1 knockout HeLa cells.

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

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.
Purification:
XBP-1 Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
XBP-1 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
XBP-1 antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
XBP1
Additional Names:
XBP-1 Antibody: XBP2, TREB5, XBP-1, XBP2, X-box-binding protein 1, Tax-responsive element-binding protein 5
Protein Accession Number:
P17861
Protein Gi Number:
60416406
Ncbi Gene Id Number:
7494
User Note:
Optimal dilutions for each application to be determined by the researcher.
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 Reference 1:
Yoshida et al. Cell 2001; 107:881-91.
Background Reference 2:
Calfon et al. Nature 2002; 415:92-6.
Background Reference 3:
Haze et al. Mol. Cell. Biol. 1999; 10:3787-99.
Background Reference 4:
Little et al. Crit. Rev. Eukaryot. Gene Expr. 1994; 4:1-18.
Citation 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
Citation 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

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

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.

CATALOG NUMBER:

3687

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

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