ATF6 Antibody (70B1413.1) - BSA Free
Novus Biologicals, part of Bio-Techne | Catalog # NBP1-40256
Conjugate
Catalog #
Forumulation
Catalog #
Key Product Details
Validated by
Tagged Protein Expression
Species Reactivity
Human, Mouse, Rat, Porcine, Plant, Rabbit
Applications
Chromatin Immunoprecipitation (ChIP), CyTOF-ready, Flow (Intracellular), Flow Cytometry, Immunoblotting, Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, Knockout Validated, Western Blot
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG1 kappa Clone # 70B1413.1
Format
BSA Free
Concentration
1.0 mg/ml
Product Specifications
Immunogen
This monoclonal antibody was made against a partial protein containing amino acids 1-273 of human ATF6.
Reactivity Notes
Plant reactivity reported in scientific literature (PMID: 31531232). Use in Plant reported in scientific literature (PMID:31531232).
Specificity
This ATF6 antibody detects both the full length and the cleaved/active protein.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG1 kappa
Description
(1) The ATF6 (IMG-273) has been cited to recognize both full-length and cleaved forms of ATF6. Please refer to the references in the Product Citation list for more comprehensive information.
(2) The ATF6 transfected cell lysate (40210) is recommended as a useful western blot positive control to run in parallel with your experimental samples.
(3) Active/cleaved forms of ATF6 are generated through proteolytic cleavage during ER stress. Different molecular weights have been described for cleaved forms. A sample protocol was first described in Luo and Lee (2002) wherein NIH3T3 cells were treated with the amino acid analogue azetidine (AzC), and full-length and cleaved forms ATF6 were detected with IMG-273. The results show that in addition to the major 90 kDa full length ATF6, protein bands over the range of 50-70 kDa were detectable following AzC treatment (Luo and Lee, 2002: Figure 4A, page 791). Many protocols and other cleaved forms have been described since, please consult the literature for additional information.
(4) For immunofluorescence microscopy, cells were fixed in methanol at -20 degrees C (Thomas et al, 2005). Thomas et al used immunofluorescence to identify ATF6 with IMG-273 in the nucleus.
(5) The active/cleaved 50 kDa nuclear form of ATF6 using IMG-273 has been found to be strongly expressed in certain tumor cell lines derived from B cell lymphoma (DEL), primary effusion lymphoma [BC-3 (ATCC CRL-2277), PEL-SY, HBL-6], lymphoblastic leukemia (DS-1) and multiple myeloma (RPMI-8226, NCI-H929), (Jenner et al. 2003).
(6) Cleaved 60 and 36 kDa ATF6 forms have also been described in the nucleus (Mao et al, 2007).
(7) The ATF6 antibody is reported to be specific for ATF6a, recognizing ATF6a but not ATF6b (Bommiasamy et al, 2009).
(8) In western blots, the binding pattern of ATF6 may vary. Researchers are encouraged to consult the body of literature citing the ATF6 IMG-273 antibody (see Product citation list) for additional information. General ATF6 literature is also helpful. For example, Yoshida (1998) show a multiple band pattern in HeLa in both untreated and stressed cells (Fig 11B). In this early landmark ATF6 publication, multiple bands were seen between the 66 and 116 kDa markers as well as one or more bands between the 45 and 66 kDa markers.
(9) We highly recommend the use of a maximum sensitivity ECL substrate (Femto sensitive) for efficient detection of this antibody in Western blot applications.
(2) The ATF6 transfected cell lysate (40210) is recommended as a useful western blot positive control to run in parallel with your experimental samples.
(3) Active/cleaved forms of ATF6 are generated through proteolytic cleavage during ER stress. Different molecular weights have been described for cleaved forms. A sample protocol was first described in Luo and Lee (2002) wherein NIH3T3 cells were treated with the amino acid analogue azetidine (AzC), and full-length and cleaved forms ATF6 were detected with IMG-273. The results show that in addition to the major 90 kDa full length ATF6, protein bands over the range of 50-70 kDa were detectable following AzC treatment (Luo and Lee, 2002: Figure 4A, page 791). Many protocols and other cleaved forms have been described since, please consult the literature for additional information.
(4) For immunofluorescence microscopy, cells were fixed in methanol at -20 degrees C (Thomas et al, 2005). Thomas et al used immunofluorescence to identify ATF6 with IMG-273 in the nucleus.
(5) The active/cleaved 50 kDa nuclear form of ATF6 using IMG-273 has been found to be strongly expressed in certain tumor cell lines derived from B cell lymphoma (DEL), primary effusion lymphoma [BC-3 (ATCC CRL-2277), PEL-SY, HBL-6], lymphoblastic leukemia (DS-1) and multiple myeloma (RPMI-8226, NCI-H929), (Jenner et al. 2003).
(6) Cleaved 60 and 36 kDa ATF6 forms have also been described in the nucleus (Mao et al, 2007).
(7) The ATF6 antibody is reported to be specific for ATF6a, recognizing ATF6a but not ATF6b (Bommiasamy et al, 2009).
(8) In western blots, the binding pattern of ATF6 may vary. Researchers are encouraged to consult the body of literature citing the ATF6 IMG-273 antibody (see Product citation list) for additional information. General ATF6 literature is also helpful. For example, Yoshida (1998) show a multiple band pattern in HeLa in both untreated and stressed cells (Fig 11B). In this early landmark ATF6 publication, multiple bands were seen between the 66 and 116 kDa markers as well as one or more bands between the 45 and 66 kDa markers.
(9) We highly recommend the use of a maximum sensitivity ECL substrate (Femto sensitive) for efficient detection of this antibody in Western blot applications.
Scientific Data Images for ATF6 Antibody (70B1413.1) - BSA Free
Western Blot: ATF6 Antibody (70B1413.1) [NBP1-40256] -
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.
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.
Immunocytochemistry/Immunofluorescence: ATF6 Antibody (70B1413.1) [NBP1-40256] - Untreated HeLa cells were fixed in -20C methanol for 10 min, air dried and rehydrated in PBS at room temperature for 5 minutes. Cells were incubated with anti-ATF6 (1:20) for one hour at room temperature. ATF6 reactivity (green) was detected with anti-mouse Dylight-488 secondary antibody. Nuclei were counterstained with DAPI (blue). Note the ER localization of ATF6.
Applications for ATF6 Antibody (70B1413.1) - BSA Free
Application
Recommended Usage
Chromatin Immunoprecipitation (ChIP)
1:10-1:500
Flow Cytometry
1 ug per million cells
Immunocytochemistry/Immunofluorescence
1:10-1:500
Immunohistochemistry
1:10-1:500
Immunohistochemistry-Frozen
1:10-1:500
Immunohistochemistry-Paraffin
1:50
Immunoprecipitation
1:10-1:500
Western Blot
1-5 ug/ml
Application Notes
ICC: See Thomas et al (2005) and Kikuchi et al (2006) for details. Immunohistochemistry (Frozen): See Zhu et al (2008) for details. Immunohistochemistry (Paraffin): See van Kollenburg et al (2006) for details. Immunoprecipitation: See Hong et al, 2004 for details. Use in Immunoblotting reported in scientific literature (PMID 28550308). Knockout validation (PMID: 31531232). In Western blot do not use milk in the diluent as it can inhibit the observed signal. This antibody is CyTOF ready.
Reviewed Applications
Read 3 reviews rated 3.7 using NBP1-40256 in the following applications:
Formulation, Preparation, and Storage
Purification
Protein G purified
Formulation
TBS
Format
BSA Free
Preservative
0.05% Sodium Azide
Concentration
1.0 mg/ml
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Background: ATF6
Long Name
Activating Transcription Factor 6
Alternate Names
ACHM7, ATF6 alpha
Entrez Gene IDs
22926 (Human)
Gene Symbol
ATF6
UniProt
Additional ATF6 Products
Product Documents for ATF6 Antibody (70B1413.1) - BSA Free
Product Specific Notices for ATF6 Antibody (70B1413.1) - BSA Free
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
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