Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸

Order now and get it on Tuesday March 02, 2021

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

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

Application	Species
IHC-P	Mouse Human
WB	Rat Human

See all applications and species data

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
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: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR14774
Isotype

IgG
Research areas

Western blot - Anti-TXNIP antibody [EPR14774] (ab188865)

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 kDa

Lanes 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.
Western blot - Anti-TXNIP antibody [EPR14774] (ab188865)

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
Western blot - Anti-TXNIP antibody [EPR14774] (ab188865)

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?
Western blot - Anti-TXNIP antibody [EPR14774] (ab188865)

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?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TXNIP antibody [EPR14774] (ab188865)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TXNIP antibody [EPR14774] (ab188865)

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.
Western blot - Anti-TXNIP antibody [EPR14774] (ab188865)

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 seconds

Blocking and diluting buffer: 5% NFDM/TBST

MG-132 induces the expression of TXNIP (PMID: 20068034). H2O2 reduces the expression of TXNIP (PMID: 24376827).
Anti-TXNIP antibody [EPR14774] (ab188865)

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