Anti-XBP1 Antibody Picoband™

Product Name

Anti-XBP1 Antibody Picoband™

View all XBP1 Antibodies

SKU/Catalog Number

PB9463

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-XBP1 Antibody Picoband™ catalog # PB9463. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-XBP1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9463)

Host

Rabbit

Contents

Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence at the N-terminus of human XBP1, identical to the related mouse and rat sequences.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins

Reactive Species

PB9463 is reactive to XBP1 in Human, Mouse, Rat

Applications

PB9463 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

29 kDa

Calculated molecular weight

28.695kDa

Background of XBP1

XBP1, also known as X-box binding protein 1, is a protein which in humans is encoded by the XBP1 gene. It encodes a transcription factor that regulates MHC class II genes by binding to a promoter element referred to as an X box. This gene product is a bZIP protein, which was also identified as a cellular transcription factor that binds to an enhancer in the promoter of the T cell leukemia virus type 1 promoter. It may increase expression of viral proteins by acting as the DNA binding partner of a viral transactivator. It has been found that upon accumulation of unfolded proteins in the endoplasmic reticulum (ER), the mRNA of this gene is processed to an active form by an unconventional splicing mechanism that is mediated by the endonuclease inositol-requiring enzyme 1 (IRE1). The resulting loss of 26 nt from the spliced mRNA causes a frame-shift and an isoform XBP1 (S), which is the functionally active transcription factor. The isoform encoded by the unspliced mRNA, XBP1 (U), is constitutively expressed, and thought to function as a negative feedback regulator of XBP1 (S), which shuts off transcription of target genes during the recovery phase of ER stress. A pseudogene of XBP1 has been identified and localized to chromosome 5.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, By Heat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of XBP using anti-XBP antibody (PB9463).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours.
Lane 1: MCF-7 Whole Cell Lysate at 40ug,
Lane 2: MM231 Whole Cell Lysate at 40ug,
Lane 3: MM453 Whole Cell Lysate at 40ug,
Lane 4: SKOV Whole Cell Lysate at 40ug,
Lane 5: HELA Whole Cell Lysate at 40ug.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-XBP antigen affinity purified polyclonal antibody (Catalog # PB9463) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for XBP at approximately 29 kDa. The expected band size for XBP is at 29 kDa.

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Figure 2. IHC analysis of XBP using anti-XBP antibody (PB9463).
XBP was detected in a paraffin-embedded section of mouse lung tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-XBP Antibody (PB9463) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 3. IHC analysis of XBP using anti-XBP antibody (PB9463).
XBP was detected in a paraffin-embedded section of rat lung tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-XBP Antibody (PB9463) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 4. IHC analysis of XBP using anti-XBP antibody (PB9463).
XBP was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-XBP Antibody (PB9463) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 5. Flow Cytometry analysis of HepG2 cells using anti-XBP1 antibody (PB9463).
Overlay histogram showing HepG2 cells stained with PB9463 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-XBP1 Antibody (PB9463, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For XBP1 (Source: Uniprot.org, NCBI)

Alternative Names

HTF; TREB5; X-box binding protein 1; X-box-binding protein 1; XBP1; XBP-1; XBP2; XBP2Tax-responsive element-binding protein 5 XBP1 TREB-5, TREB5, XBP-1, XBP2 X-box binding protein 1 X-box-binding protein 1|tax-responsive element-binding protein 5

For more info on XBP1, check out the XBP1 Infographic

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In this infographic, you will see the following information for XBP1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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