Anti-TXNIP antibody [EPR14774] (ab188865)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR14774] to TXNIP
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
-
Product name
Anti-TXNIP antibody [EPR14774]
See all TXNIP primary antibodies -
Description
Rabbit monoclonal [EPR14774] to TXNIP -
Host species
Rabbit -
Specificity
Expression levels of the target protein vary with sample type and some optimisation may be required. For western blotting, more concentrated lysates may be required when using tissues samples.
Stimulation may be required to allow detection of the target protein due to low levels of endogenous expression in some samples. Please see images below for recommended treatment conditions and positive controls.
-
Tested Applications & Species
See all applications and species dataApplication Species IHC-P MouseHumanWB RatHuman -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- IHC-P: Human kidney and Mouse liver tissue. WB: HeLa, 293T, HAP1, MCF7, K562, THP-1, MDA-MB-231, BxPC-3, HUVEC, BxPC-3, PC-12 and NIH/3T3; Human liver, skeletal muscle and kidney tissue lysates
-
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
-
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: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR14774 -
Isotype
IgG -
Research areas
Images
-
All lanes : Anti-TXNIP antibody [EPR14774] (ab188865) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : TXNIP knockout HAP1 whole cell lysate
Lane 3 : MCF7 whole cell lysate
Lane 4 : K562 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 44 kDa
Observed band size: 44 kDaLanes 1 - 4: Merged signal (red and green). Green - ab188865 observed at 44 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab188865 was shown to recognize TXNIP in wild-type HAP1 cells as signal was lost at the expected MW in TXNIP knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and TXNIP knockout samples were subjected to SDS-PAGE. ab188865 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
-
All lanes : Anti-TXNIP antibody [EPR14774] (ab188865) at 1/1000 dilution
Lane 1 : THP-1 (human acute monocytic leukemia) whole cell lysates
Lane 2 : MCF7 (human breast carcinoma) whole cell lysates
Lane 3 : MDA-MB-231 (human mammary gland epithelial adenocarcinoma) whole cell lysates
Lane 4 : BxPC-3 (human Pancreas adenocarcinoma) whole cell lysates
Lane 5 : HUVEC (human umbilical vein/vascular endothelium) whole cell lysates (negative/low expression)
Lane 6 : Human liver lysate
Lane 7 : Human kidney lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 44 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?Blocking buffer and concentration 5% NFDM/TBST
Diluting buffer and concentration 5% NFDM/TBST -
All lanes : Anti-TXNIP antibody [EPR14774] (ab188865) at 1/5000 dilution
Lane 1 : Human skeletal muscle lysate
Lane 2 : BxPC-3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution
Predicted band size: 44 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?
-
All lanes : Anti-TXNIP antibody [EPR14774] (ab188865) at 1/1000 dilution
Lane 1 : PC-12 cell lysate
Lane 2 : NIH/3T3 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution
Predicted band size: 44 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?
-
Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling TXNIP with ab188865 at 1/250 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling TXNIP with ab188865 at 1/250 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
All lanes : Anti-TXNIP antibody [EPR14774] (ab188865) at 1/1000 dilution
Lane 1 : 293T (Human embryonic kidney epithelial cell) whole cell lysates
Lane 2 : 293T (Human embryonic kidney epithelial cell) treated with 25uM MG-132 for 4 hours whole cell lysates.
Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 4 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 10mM H2O2 for 1 hour whole cell lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 44 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking and diluting buffer: 5% NFDM/TBST
MG-132 induces the expression of TXNIP (PMID: 20068034). H2O2 reduces the expression of TXNIP (PMID: 24376827).
-