HIF1-alpha Antibody
Rabbit Polyclonal
Antigen Affinity Purified

Protein ID

Q16665

Catalog No.

A300-286A

GeneID

3091

Lot No.

A300-286A-7

APPLICATIONSWB, IP, IHC, ICC-IF

SPECIES REACTIVITY

Human, Mouse

PRESUMED REACTIVITYAMOUNT

Based on 100% sequence identity, this antibody is predicted to react with Rat, Chicken andBovine
100 µlCONCENTRATION

200 µg/ml

STORAGE/SHELF LIFE

2 - 8°C / 1 year from date of receipt

PHYSICAL STATE

LiquidBUFFER

Tris-buffered Saline containing 0.1% BSA and 0.09% Sodium Azide

ISOTYPE

IgG

ORIGINUSAPRODUCTIONPROCEDURES

Antibody was affinity purified using an epitope specific to HIF1-alpha immobilized on solidsupport.
The epitope recognized by A300-286A maps to a region between residues 775 and the Cterminus (residue 826) of human hypoxia-inducible factor 1, alpha subunit using thenumbering given in Swiss-Prot entry Q16665 (GeneID 3091).
Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280
nm of 1.4 equals 1.0 mg of IgG.

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

1:2,000 - 1:10,000

Immunoprecipitation

2 - 10 µg/mg lysate

Immunohistochemistry

1:100 to 1:500. Epitope retrieval with citrate buffer pH6.0 for 20
minutes using a pressure cooker is recommended for FFPE tissue
sections.

Immunofluorescence(ICC)

1:100 - 1:500. Epitope retrieval with citrate buffer pH 6.0 for 20
minutes using a pressure cooker is recommended for FFPE tissue
sections.

IHC HUMAN CONTROLSRenal Cell Carcinoma, HeLa Cells, HeLa Cells Hypoxic

ADDITIONAL INFO

https://www.bethyl.com/product/A300-286A
Use the link above to view SDS, a current list of citations, and other product specific information.
IP-western blot protocol: https://www.bethyl.com/content/protocol_IP_WB

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

Date: April 14, 2022

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

1For 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.

HIF1-alpha AntibodyA300-286A

Detection of human HIF1 alpha by western blot. Samples:
Whole cell lysate (50 µg) from HepG2 cells that were either
treated with 200 µM cobalt chloride (+) or mock treated(-). Antibodies: Affinity purified rabbit anti-HIF1 alphaantibody A300-286A (lot number A300-286A-7) used for
WB at 0.04 µg/ml. Detection: Chemiluminescence with
exposure times of 10 seconds. Lower Panel: Rabbit antiCOPB2 antibody (A304-523A).

Detection of mouse HIF1 alpha by western blot. Samples:
Whole cell lysate (50 µg) from NIH3T33ells that were either
treated with 200 µM cobalt chloride (+) or mock treated(-). Antibodies: Affinity purified rabbit anti-HIF1 alphaantibody A300-286A (lot number A300-286A-7) used for
WB at 0.04 µg/ml. Detection: Chemiluminescence with
exposure times of 3 seconds. Lower Panel: Rabbit antiCOPB2 antibody (A304-523A).

Detection of human HIF1 alpha by western blot of
immunoprecipitates. Samples: Whole cell lysate (1 mg for
IP, 20% of IP loaded) from HepG2 cells that were treatedwith 200 µM cobalt chloride or untreated. Antibodies:Affinity purified rabbit anti-HIF1 alpha antibody
A300-286A (lot number A300-286A-7) used for WB at
0.04 µg/ml and for IP at 6 µg/mg lysate. Detection:
Chemiluminescence with exposure times of 3 seconds.

Detection of human HIF1-alpha byimmunohistochemistry. Sample: FFPE section of renal cellcarcinoma. Antibody: Affinity purified rabbit anti-HIF1alpha antibody (Cat. No. A300-286A Lot6) used at adilution of 1:100 (2µg/ml). Detection: DAB

Phone: 800.338.9579 • Fax: 866.597.6105 • Web: www.bethyl.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.

HIF1-alpha Antibody

A300-286A

Detection of human HIF1-alpha by immunocytochemistry.
Sample: Formaldehyde-fixed asynchronous HeLa cells
treated with 200 µM cobalt chloride (left) or mocktreatment (right). Antibody: Affinity purified rabbit antiHIF1-alpha antibody Cat. No. A300-286A Lot6 (upperimages) and A300-286A Lot5 (lower images) used at adilution of 1:100 (2µg/ml). Detection: Red-fluorescentgoat anti-rabbit IgG-heavy and light chain cross-adsorbed
Antibody DyLight® 594 Conjugated (A120-201D4).
Counterstain: DAPI (blue) and fluorescent phalloidin(green).

Phone: 800.338.9579 • Fax: 866.597.6105 • Web: www.bethyl.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.