Antibodies, Proteins, Kits and Reagents for Life Science

304 886 ₸ Product size

100 µl 304 886 ₸

Order now and get it on Tuesday March 09, 2021

Anti-XBP1 antibody [EPR22004] (ab220783)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR22004] to XBP1
Suitable for: Flow Cyt, IP, WB
Reacts with: Mouse, Human

Product name

Anti-XBP1 antibody [EPR22004]
See all XBP1 primary antibodies
Description

Rabbit monoclonal [EPR22004] to XBP1
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
IP	Human
WB	Mouse Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Huh7 treated with 300 nM thapsigargin for 18 hours, whole cell lysate; RAW 264.7 treated with 300 nM thapsigargin for 18 hours, whole cell lysate; U266B1, MCF7, HEK-293T and NIH/3T3 whole cell lysates. Flow Cyt: Huh7 treated with 300 nM Thapsigargin for 18 hours. IP: Huh7 treated with 300 nM Thapsigargin for 18 hours whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR22004
Isotype

IgG
Research areas

Western blot - Anti-XBP1 antibody [EPR22004] (ab220783)

All lanes : Anti-XBP1 antibody [EPR22004] (ab220783) at 1/1000 dilution

Lane 1 : Untreated Huh7 (human hepatocellular carcinoma epithelial cell line) whole cell lysate
Lane 2 : Huh7 treated with 300 nM thapsigargin for 18 hours, whole cell lysate
Lane 3 : Untreated RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 4 : RAW 264.7 treated with 300 nM thapsigargin for 18 hours, whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 29 kDa
Observed band size: 29,56 kDa
why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

Spliced XBP1 (XBP1s) is induced after ER stress which is mimicked by treatment with thapsigargin. The expression profile observed is consistent with what has been described in the literature (PMID:11779464; PMID 25190803).
Flow Cytometry - Anti-XBP1 antibody [EPR22004] (ab220783)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Huh7 (human hepatocellular carcinoma epithelial cell line) cells treated with 300 nM Thapsigargin for 18 hours (red) / Untreated control (green) labeling XBP1 with ab220783 at 1/100 dilution compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells incubated with secondary antibody only) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-XBP1 antibody [EPR22004] (ab220783)

XBP1 was immunoprecipitated from 0.35 mg Huh7 (human hepatocellular carcinoma epithelial cell line) treated with 300 nM Thapsigargin for 18 hours whole cell lysate with ab220783 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab220783 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1: Huh7 treated with 300 nM Thapsigargin for 18 hours whole cell lysate 10 µg (Input).
Lane 2: ab220783 IP in Huh7 treated with 300 nM Thapsigargin for 18 hours whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab220783 in Huh7 treated with 300 nM Thapsigargin for 18 hours whole cell lysate (-).

Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
Western blot - Anti-XBP1 antibody [EPR22004] (ab220783)

All lanes : Anti-XBP1 antibody [EPR22004] (ab220783) at 1/1000 dilution

Lane 1 : U266B1 (human multiple myeloma B lymphocyte cell line) whole cell lysate
Lane 2 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
Lane 3 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 4 : NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 29 kDa
Observed band size: 29,56 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The expression profile observed is consistent with what has been described in the literature (PMID: 11779464; PMID 25190803).
Anti-XBP1 antibody [EPR22004] (ab220783)

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