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[KO Validated] HIF1AN/FIH-1 Rabbit mAb (A27299)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse, Rat

Western blot analysis of various lysates using [KO Validated] HIF1AN/FIH-1 Rabbit mAb (A27299) at 1:16000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 45s.

Western blot analysis of lysates from wild type (WT) and HIF1AN/FIH-1 knockout (KO) 293T cells using [KO Validated] HIF1AN/FIH-1 Rabbit mAb (A27299) at 1:16000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 45s.

ABclonal:Immunohistochemistry - [KO Validated] HIF1AN/FIH-1 Rabbit mAb (A27299)

Immunohistochemistry analysis of paraffin-embedded Human colon tissue using [KO Validated] HIF1AN/FIH-1 Rabbit mAb (A27299) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human esophagus tissue using [KO Validated] HIF1AN/FIH-1 Rabbit mAb (A27299) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse colon tissue using [KO Validated] HIF1AN/FIH-1 Rabbit mAb (A27299) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat testis tissue using [KO Validated] HIF1AN/FIH-1 Rabbit mAb (A27299) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded 293T and 293T-HIF1AN-KO cells using [KO Validated] HIF1AN/FIH-1 Rabbit mAb (A27299) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

ABclonal:Immunofluorescence - [KO Validated] HIF1AN/FIH-1 Rabbit mAb (A27299)

Confocal imaging of NIH/3T3 cells using [KO Validated] HIF1AN/FIH-1 Rabbit mAb (A27299, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

Overview

Product name	[KO Validated] HIF1AN/FIH-1 Rabbit mAb
Catalog No.	A27299
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC73051

Background

Enables several functions, including 2-oxoglutarate-dependent dioxygenase activity; NF-kappaB binding activity; and transition metal ion binding activity. Involved in several processes, including negative regulation of Notch signaling pathway; negative regulation of transcription from RNA polymerase II promoter in response to hypoxia; and protein hydroxylation. Located in cytosol; nucleoplasm; and perinuclear region of cytoplasm. Colocalizes with nucleus.

Immunogen information

Immunogen	Recombinant fusion protein containing a sequence corresponding to amino acids 1-349 of human HIF1AN/FIH1 (NP_060372.2).
Sequence	MAATAAEAVASGSGEPREEAGALGPAWDESQLRSYSFPTRPIPRLSQSDPRAEELIENEEPVVLTDTNLVYPALKWDLEYLQENIGNGDFSVYSASTHKFLYYDEKKMANFQNFKPRSNREEMKFHEFVEKLQDIQQRGGEERLYLQQTLNDTVGRKIVMDFLGFNWNWINKQQGKRGWGQLTSNLLLIGMEGNVTPAHYDEQQNFFAQIKGYKRCILFPPDQFECLYPYPVHHPCDRQSQVDFDNPDYERFPNFQNVVGYETVVGPGDVLYIPMYWWHHIESLLNGGITITVNFWYKGAPTPKRIEYPLKAHQKVAIMRNIEKMLGEALGNPQEVGPLLNTMIKGRYN
Gene ID
Swiss Prot
Synonyms	FIH1
Calculated MW	41kDa
Observed MW	40kDa

Applications

Reactivity	Human, Mouse, Rat
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:8000 - 1:32000 IHC-P 1:100 - 1:400 IF/ICC 1:100 - 1:400 ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.09% Sodium azide, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunofluorescence
Positive samples	293T, HeLa, NIH/3T3, PC-12, Mouse brain, Rat brain
Cellular location	Cytoplasm, Nucleus, perinuclear region.