NBP1-40256-0.1mg

antibody from Novus Biologicals

Targeting: ATF6 ATF6A

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Data presented on provider website)

Immunoprecipitation (Data presented on provider website)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Data presented on provider website)

Antibody Data

Product number

NBP1-40256-0.1mg

Provider

Novus Biologicals

Proper citation

Novus Cat#NBP1-40256-0.1mg, RRID:AB_11005857

Product name

Mouse Monoclonal ATF6 Antibody

Antibody type

Monoclonal

Description

Protein G purified. This ATF6 antibody detects both the full length and the cleaved/active protein.

Reactivity

Human, Mouse, Rat, Porcine, Rabbit

Host

Mouse

Isotype

IgG

Vial size

0.1 mg

Concentration

1.0 mg/ml

Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Western blot

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

Western Blot: ATF6 Antibody (70B1413.1) [NBP1-40256] - Lane 1: 293 cells transfected with full-length ATF6. Lane 2: 293 cells transfected with partial length ATF6 (amino acids 1-373). Lane 3: Untransfected 293 cells. Western blots were probed with 4 ug/ml of the ATF6 monoclonal antibody and visualized with PicoTect Western Blot Chemiluminescence Substrate (10087K). Film was exposed for 1 min. The top arrow corresponds to the 90 kDa form of ATF6 described as full-length in the literature. The human full-length and partial length ATF6 plasmids are described in Luo and Lee (2002).

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

Western Blot: ATF6 Antibody (70B1413.1) [NBP1-40256] - Analysis of ATF6 in mouse liver tissue using 3 ug/ml of ATF6 antibody and 0.25 ug/ml of GAPDH antibody. Lane A contains 20 ugs of whole mouse liver lysate, lane B contains 20 ugs of total ER fraction, and lane C contains 20 ugs of rough ER fraction. The ATF6 band may represent under glycosylated or cleaved/active ATF6.

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

Western Blot: ATF6 Antibody (70B1413.1) [NBP1-40256] - Analysis of ATF6 in NIH3T3 cell lysate using NBP1-40256 at 3 ug/ml. A band corresponding to full-length ATF6 was detected. We have not characterized the ~36 kDa observed band; it may be an ATF6 breakdown/cleavage product.

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

Western Blot: ATF6 Antibody (70B1413.1) [NBP1-40256] - Image of anti-ATF6. Whole cell protein from Hek293 and HepG2 treated with and without 10 mM DTT was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 5 ug/ml anti-ATF6 and detected with an anti-mouse HRP secondary antibody using a maximum sensitivity ECL reagent. Note the loss of the 90 kDa modified ATF6 protein (arrowhead) upon DTT treatment to activate the unfolded protein response.

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

Western Blot: ATF6 Antibody (70B1413.1) [NBP1-40256] - Image of anti-ATF6. Whole cell protein from HeLa cells treated with and without 10 mM DTT was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 5 ug/ml anti-ATF6 and detected with an anti-mouse HRP secondary antibody using a maximum sensitivity ECL reagent. Note the loss of the 90 kDa modified ATF6 protein (arrowhead) upon DTT treatment to activate the unfolded protein response.

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

Western Blot: ATF6 Antibody (70B1413.1) [NBP1-40256] - Hypoxia leads to EMT and ER-stress in CRC cells. B. C. Confluent growing SW480 (B) and HCT116 (C) cells were cultured under conditions of normoxia or hypoxia-like conditions (serum free; 100 uM CoCl2, 1-9 h. B-actin (B-act) as loading control. HIF1a was detectable after 3 h of CoCl2 incubation, the amount of the 50 kD-ATF6 fragment, was already enhanced after 1 h of addition of CoCl2. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0087386) licensed under a CC-BY licence.

Immunohistochemistry

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

Immunohistochemistry-Paraffin: ATF6 Antibody (70B1413.1) [NBP1-40256] - Human placenta, followed by biotinylated horse anti-mouse IgG secondary antibody, alkaline phosphatase-streptavidin and chromogen. Dilution 10ug/ml

Flow cytometry

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

Flow Cytometry: ATF6 Antibody (70B1413.1) [NBP1-40256] - Intracellular flow cytometric staining of 1 x 10^6 MCF-7 cells using ATF6 antibody (dark blue). Isotype control shown in orange. An antibody concentration of 1 ug/1x10^6 cells was used.

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

Flow (Intracellular): ATF6 Antibody (70B1413.1) [NBP1-40256] - An intracellular stain was performed on HeLa cells with ATF6 Antibody (70B1413.1) NBP1-40256F (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to FITC.