Human/Mouse/Rat ATF6 Antibody

R&D Systems | Catalog # MAB71527

Recombinant Monoclonal Antibody.
R&D Systems
Discontinued Product
MAB71527 has been discontinued. An alternative/replacement product is available: NBP1-75478. View all ATF6 products.

Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human

Applications

Validated:

Immunohistochemistry, Western Blot, Intracellular Staining by Flow Cytometry, Immunocytochemistry

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 2358C
View all ATF6 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human ATF6
Met1-Thr192
Accession # P18850

Specificity

Detects human ATF6 in direct ELISAs. Detects human, mouse, and rat ATF6 in Western blots.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat ATF6 Antibody

Detection of Human, Mouse, and Rat ATF6 antibody by Western Blot.

Detection of Human, Mouse, and Rat ATF6 by Western Blot.

Western blot shows lysates of Raji human Burkitt's lymphoma cell line, Jurkat human acute T cell leukemia cell line, A20 mouse B cell lymphoma cell line, and PC-12 rat adrenal pheochromocytoma cell line. PVDF membrane was probed with 0.05 µg/mL of Rabbit Anti-Human/Mouse/Rat ATF6 Monoclonal Antibody (Catalog # MAB71527) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for ATF6 at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
ATF6 antibody in Human Prostate by Immunohistochemistry (IHC-P).

ATF6 in Human Prostate.

ATF6 was detected in immersion fixed paraffin-embedded sections of human prostate using Rabbit Anti-Human/Mouse/Rat ATF6 Monoclonal Antibody (Catalog # MAB71527) at 0.3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
ATF6 antibody in A549 Human Cell Line by Immunocytochemistry (ICC).

ATF6 in A549 Human Cell Line.

ATF6 was detected in immersion fixed A549 human lung carcinoma cell line using Rabbit Anti-Human/Mouse/Rat ATF6 Monoclonal Antibody (Catalog # MAB71527) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of ATF6 antibody in Human HeLa cells antibody by Flow Cytometry.

Detection of ATF6 in Human HeLa cells by Flow Cytometry.

HeLa human cervical epithelial cell line was stained with Rabbit Anti-Human/Mouse/Rat ATF6 Monoclonal Antibody (Catalog # MAB71527, filled histogram) or Rabbit IgG control antibody (Catalog # MAB1050, open histogram) followed by APC-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0111). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.
Detection of ATF6 antibody in Human MCF-7 cells antibody by Flow Cytometry.

Detection of ATF6 in Human MCF-7 cells by Flow Cytometry.

MCF-7 human breast cancer cell line was stained with Rabbit Anti-Human/Mouse/Rat ATF6 Monoclonal Antibody (Catalog # MAB71527, filled histogram) or Rabbit IgG control antibody (Catalog # MAB1050, open histogram) followed by APC-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0111). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.

Applications for Human/Mouse/Rat ATF6 Antibody

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed A549 human lung carcinoma cell line

Immunohistochemistry

0.3-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human prostate

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: HeLa human cervical epithelial cell line and MCF-7 human breast cancer cell line, fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012).

Western Blot

0.05 µg/mL
Sample: Raji human Burkitt's lymphoma cell line, Jurkat human acute T cell leukemia cell line, A20 mouse B cell lymphoma cell line, and PC‑12 rat adrenal pheochromocytoma cell line

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: ATF6

ATF6 is a constitutively expressed, endoplasmic reticulum (ER) membrane-anchored transcription factor. ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to environmental and physiological stresses that target the ER (reviewed in Shen, 2005 & Prywes, 2005). The C-terminus of ATF6 is located in the ER lumen and its N-terminal DNA binding domain faces the cytosol. AFT6 plays a key role in the ER stress response by transmitting the ER stress signal across the ER membrane into the nucleus. The induction of new gene expression by ATF6 is an important aspect of the ER stress response. In response to certain stress conditions, ATF6 translocates from the ER to the Golgi. The 90 kDa full-length ATF6 is processed within the Golgi to its active 50 kDa form through sequential cleavage by site-1 and site-2 proteases (S1P and S2P). Proteolytic activation of ATF6 in the ER stress response is a mechanism to regulate membrane-bound factors, and is referred to as regulated intramembrane proteolysis. The N-terminal active ATF6 translocates to the nucleus where it binds to ER stress-response elements in ER stress-response genes (ERSRGs). ATF6 is a potent transcriptional activator of ERSRGs. The fully glycosylated form of ATF6, a 670 amino acid protein, exhibits an electrophoretic mobility of ~90 kDa in denaturing SDS-gels, in part because of the glycosylated modifications. ATF6 has 3 consensus sites for N-linked glycosylation and exists constitutively as a glycosylated protein. Differentially glycosylated ATF6 forms may result from mutations or experimental treatment.

Long Name

Activating Transcription Factor 6

Alternate Names

ACHM7, ATF6 alpha

Entrez Gene IDs

22926 (Human); 226641 (Mouse); 304962 (Rat)

Gene Symbol

ATF6

UniProt

P18850

Additional ATF6 Products

Product Documents for Human/Mouse/Rat ATF6 Antibody

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Product Specific Notices for Human/Mouse/Rat ATF6 Antibody

For research use only

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Protocols

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