Anti-TXNIP antibody [EPR14774]

• IHC-P

Unknown

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.

• IHC-P

Supplier Data

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.

• WB

Supplier Data

Western blot - Anti-TXNIP antibody [EPR14774] (AB188865)

All lanes:

Western blot - Anti-TXNIP antibody [EPR14774] (ab188865) at 1/5000 dilution

Lane 1:

Human skeletal muscle lysate at 20 µg

Lane 2:

BxPC-3 cell lysate at 20 µg

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

false

• WB

Lab

Western blot - Anti-TXNIP antibody [EPR14774] (AB188865)

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.

All lanes:

Western blot - Anti-TXNIP antibody [EPR14774] (ab188865) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

TXNIP knockout HAP1 whole cell lysate at 20 µg

Lane 3:

MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

K-562 (human chronic myelogenous leukemia lymphoblasts cell line) whole cell lysate at 20 µg

Predicted band size: 44 kDa

Observed band size: 44 kDa

false

• WB

Lab

Western blot - Anti-TXNIP antibody [EPR14774] (AB188865)

Blocking buffer and concentration 5% NFDM/TBST
Diluting buffer and concentration 5% NFDM/TBST

All lanes:

Western blot - Anti-TXNIP antibody [EPR14774] (ab188865) at 1/1000 dilution

Lane 1:

THP-1 (human acute monocytic leukemia) whole cell lysates at 20 µg

Lane 2:

MCF7 (human breast carcinoma) whole cell lysates at 20 µg

Lane 3:

MDA-MB-231 (human mammary gland epithelial adenocarcinoma) whole cell lysates at 20 µg

Lane 4:

BxPC-3 (human Pancreas adenocarcinoma) whole cell lysates at 20 µg

Lane 5:

HUVEC (human umbilical vein/vascular endothelium) whole cell lysates (negative/low expression) at 20 µg

Lane 6:

Human liver lysate at 20 µg

Lane 7:

Human kidney lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 44 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-TXNIP antibody [EPR14774] (AB188865)

Blocking and diluting buffer : 5% NFDM/TBST

MG-132 induces the expression of TXNIP (PMID : 20068034). H2O2 reduces the expression of TXNIP (PMID : 24376827).

All lanes:

Western blot - Anti-TXNIP antibody [EPR14774] (ab188865) at 1/1000 dilution

Lane 1:

293T (Human embryonic kidney epithelial cell) whole cell lysates at 15 µg

Lane 2:

293T (Human embryonic kidney epithelial cell) treated with 25uM MG-132 for 4 hours whole cell lysates. at 15 µg

Lane 3:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Lane 4:

HeLa (Human cervix adenocarcinoma epithelial cell) treated with 10mM H2O2 for 1 hour whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 44 kDa

Observed band size: 50 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-TXNIP antibody [EPR14774] (AB188865)

All lanes:

Western blot - Anti-TXNIP antibody [EPR14774] (ab188865) at 1/1000 dilution

Lane 1:

PC-12 cell lysate at 10 µg

Lane 2:

NIH/3T3 cell lysate at 10 µg

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

false