HIF2-alpha Recombinant Monoclonal Antibody [BL95-1A2]
Rabbit Recombinant Monoclonal
Purified
Catalog No.

A700-003-T

Lot No.

A700-003-T-3

Protein ID

NP_001421.2

GeneID2034

APPLICATIONSWB, IP, IHC, ICC, ICC-IF, ChIP-Seq

SPECIES REACTIVITY

Human

AMOUNT

20 µl (2 blots)

CONCENTRATION

400 µg/ml

STORAGE/SHELF LIFE

2 - 8°C / 1 year from date of receipt

PHYSICAL STATE

LiquidBUFFER

Borate Buffered Saline (BBS) pH 8.2 with 0.1% BSA and 0.09% Sodium Azide

ISOTYPE

IgG

CLONE #BL-95-1A2

ORIGINUSA

PRODUCTIONPROCEDURES

Recombinant antibody was purified from cell culture supernatant.

APPLICATIONSCentrifuge tube to remove product from lid. Optimal working dilutions should be determined
experimentally by the investigator. Prepare working dilution immediately before use.

The epitope recognized by A700-003-T maps to a region between residues 400 and 450 ofhuman endothelial PAS domain protein 1 using the numbering given in entry NP_001421.2(GeneID 2034).

Western Blot

1:1000Immunoprecipitation

20 µl/mg lysate

Immunohistochemistry

1:200 - 1:1,000. Epitope retrieval with citrate buffer pH 6.0 for 20
minutes using a pressure cooker is recommended for FFPE tissue
sections. Overnight incubations are suggested.

Immunocytochemistry

1:200 - 1:1,000. Formaldehyde fixation is recommended.
Permeabilization with Triton-X 100 is recommended forformaldehyde-fixed cells.

Immunofluorescence(ICC)

1:100 - 1:500. Formaldehyde fixation is recommended.
Permeabilization with Triton-X 100 is recommended forformaldehyde-fixed cells.

ChIP-Seq

A previous lot has qualified for ChIP-Seq. 10 - 40 µl per 30 µg
chromatin.

IHC HUMAN CONTROLS

Renal Cell Carcinoma, 786-O Cells, Hep-G2 Cells

ADDITIONAL INFO

https://www.fortislife.com/p/A700-003-T
Use the link above to view SDS, a current list of citations, and other product specific information.

This document certifies that this product has met all of the quality control standards defined by Bethyl Laboratories, Inc.
Michael Spencer, PhD

Phone: 800.338.9579 • Fax: 866.597.6105 • Web: www.fortislife.com
Orders: orders@fortislife.com • Support: technical@fortislife.com

Date: August 5, 2022

1

For in vitro laboratory use only. Not for any clinical, therapeutic or diagnostic use in humans or animals. Not for human or animal consumption. This product may not be resold or modified for resale without the prior written
approval of Bethyl Laboratories, Inc. The information provided in this data sheet is believed to be correct but does not purport to be all-inclusive and is intended to be used as a guide. Bethyl Laboratories, Inc. shall not beliable or responsible in any way for use of either this information or the material supplied. Disposal of hazardous material may be subject to federal, state or local laws or regulations.

HIF2-alpha Recombinant Monoclonal Antibody [BL-95-1A2]

A700-003-T

Detection of human HIF2-alpha by western blot. Samples:
Whole cell lysate (50 µg) from HepG2 and 786-O cells
treated with 200 µM CoCl2 (+) or mock treated (-).
Antibody: Rabbit anti-HIF2-alpha recombinant monoclonal
antibody [BL-95-1A2] (A700-003-T lot 3) used at 1:1000.
Secondary: HRP-conjugated goat anti-rabbit IgG(A120-101P). Chemiluminescence with an exposure timeof 75 seconds.

Detection of human HIF2-alpha by western blot ofimmunoprecipitates. Samples: Whole cell lysate (1.0 mg
per IP reaction; 20% of IP loaded) from Hep-G2 cells with
200 µM CoCl2 ( +) or mock treated (-). Antibodies: Rabbit
anti-HIF2-alpha recombinant monoclonal antibody[BL-95-1A2] (A700-003-T lot 3) used for IP at 20 µl/mglysate. HIF2-alpha was also immunoprecipitated by aprevious lot of this antibody (A700-003-T-2). For blotting
immunoprecipitated HIF2-alpha, A700-003-T was used at
1:1000. Chemiluminescence with an exposure time of 75
seconds.

Detection of human HIF2-alpha in FFPE renal cell
carcinoma by IHC. Antibody: Rabbit anti-HIF2-alpharecombinant monoclonal antibody [BL95-1A2] (A700-003T lot 3). Secondary: HRP-conjugated goat anti-rabbit IgG(A120-501P). Substrate: DAB.

Detection of human HIF2-alpha in FFPE 786-O cells byICC. Antibody: Rabbit anti-HIF2-alpha recombinantmonoclonal antibody [BL95-1A2] (A700-003-T lot 3).
Secondary: HRP-conjugated goat anti-rabbit IgG(A120-501P). Substrate: DAB.

Phone: 800.338.9579 • Fax: 866.597.6105 • Web: www.fortislife.com
Orders: orders@fortislife.com • Support: technical@fortislife.com

2For in vitro laboratory use only. Not for any clinical, therapeutic or diagnostic use in humans or animals. Not for human or animal consumption. This product may not be resold or modified for resale without the prior written
approval of Bethyl Laboratories, Inc. The information provided in this data sheet is believed to be correct but does not purport to be all-inclusive and is intended to be used as a guide. Bethyl Laboratories, Inc. shall not beliable or responsible in any way for use of either this information or the material supplied. Disposal of hazardous material may be subject to federal, state or local laws or regulations.

HIF2-alpha Recombinant Monoclonal Antibody [BL-95-1A2]

Detection of human HIF2-alpha in FFPE Hep-G2 cells with(left) and without (right) cobalt treatment by ICC.
Antibody: Rabbit anti-HIF2-alpha recombinant monoclonal
antibody [BL95-1A2] (A700-003-T lot 3). Secondary: HRPconjugated goat anti-rabbit IgG (A120-501P). Substrate:DAB.

A700-003-T

Detection of human HIF2-alpha by immunocytochemistry.
Sample: Formaldehyde-fixed Hep-G2 cells untreated (left)
and treated with CoCl2 (right). Antibody: Rabbit anti-HIF2alpha recombinant monoclonal antibody [BL-95-1A2](A700-003-T lot 2) used at 1:100. Secodnasry: DyLight®
594-conjugated goat anti-rabbit IgG (A120-201D4) used
at 1:100. Counterstain: Phalloidin Alexa Fluor® 488
conjugated (green).

Localization of HIF2-alpha Binding Sites by ChIP-sequencing. Chromatin from subcutaneous humantumor 786-O cells was immunoprecipitated with anti-HIF2-alpha antibody A700-003 and analyzed byDNA sequencing. The figure illustrates the peak distribution of HIF2-alpha binding within a 250 Kb
region of chromosome 11 as detected using anti-HIF2-alpha antibody A700-003. ChIP-seq validation
performed by Active Motif, Carlsbad, CA.

Phone: 800.338.9579 • Fax: 866.597.6105 • Web: www.fortislife.com
Orders: orders@fortislife.com • Support: technical@fortislife.com

3For in vitro laboratory use only. Not for any clinical, therapeutic or diagnostic use in humans or animals. Not for human or animal consumption. This product may not be resold or modified for resale without the prior written
approval of Bethyl Laboratories, Inc. The information provided in this data sheet is believed to be correct but does not purport to be all-inclusive and is intended to be used as a guide. Bethyl Laboratories, Inc. shall not beliable or responsible in any way for use of either this information or the material supplied. Disposal of hazardous material may be subject to federal, state or local laws or regulations.