Validation Data Gallery

Filter: AllWBIPIHCIF/ICC

WB analysis using 12658-1-AP
Various lysates were subjected to SDS PAGE followed by western blot with 12658-1-AP (P4HA1 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.
WB analysis of NIH/3T3 using 12658-1-AP
NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 12658-1-AP (P4HA1 Antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.
WB analysis of human heart using 12658-1-AP
human heart tissue were subjected to SDS PAGE followed by western blot with 12658-1-AP (P4HA1 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.
WB analysis of rat skin using 12658-1-AP
rat skin tissue were subjected to SDS PAGE followed by western blot with 12658-1-AP (P4HA1 Antibody) at dilution of 1:1500 incubated at room temperature for 1.5 hours.
WB analysis of HT-1080 using 12658-1-AP
HT-1080 cells were subjected to SDS PAGE followed by western blot with 12658-1-AP (P4HA1 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.
IP experiment of A431 using 12658-1-AP
IP result of anti-P4HA1 (IP:12658-1-AP, 3ug; Detection:12658-1-AP 1:600) with A431 cells lysate 3300ug.
IHC staining of human pancreas cancer using 12658-1-AP
Immunohistochemical analysis of paraffin-embedded human pancreas cancer tissue slide using 12658-1-AP (P4HA1 antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of human pancreas cancer using 12658-1-AP
Immunohistochemical analysis of paraffin-embedded human pancreas cancer tissue slide using 12658-1-AP (P4HA1 antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of human breast cancer using 12658-1-AP
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 12658-1-AP (P4HA1 Antibody) at dilution of 1:50 (under 10x lens).
IHC staining of human breast cancer using 12658-1-AP
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 12658-1-AP (P4HA1 Antibody) at dilution of 1:50 (under 40x lens).
IF Staining of U2OS using 12658-1-AP
Immunofluorescent analysis of (-20°C Ethanol) fixed U2OS cells using P4HA1 antibody (12658-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-phalloidin (red).

Featured Product
KD/KO Validated

P4HA1 Polyclonal antibody

P4HA1 Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Cat No. 12658-1-AP

Publications(27)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, IP, ELISA

Synonyms

EC:1.14.11.2, P4HA, Procollagen-proline,2-oxoglutarate-4-dioxygenase subunit alpha-1, Prolyl 4-hydroxylase subunit alpha-1

Formulation: PBS, Azide, Glycerol

PBS, Azide, Glycerol

Conjugate: Unconjugated

Unconjugated

Size/Concentration:

-/ -

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Overview

Tested Applications
Product Information
Publications (27)
Protocols

データシート［PDF］を印刷

MSDS

Tested Applications

Positive WB detected in	A431 cells, NIH/3T3 cells, human heart tissue, rat skin tissue, HT-1080 cells, BGC-823 cells, PC-3 cells
Positive IP detected in	A431 cells
Positive IHC detected in	human pancreas cancer tissue, human breast cancer tissue **Note: suggested antigen retrieval with TE buffer pH 9.0; () Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0***
Positive IF/ICC detected in	U2OS cells

Recommended dilution

Application	Dilution
Western Blot (WB)	WB : 1:2000-1:10000
Immunoprecipitation (IP)	IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
Immunohistochemistry (IHC)	IHC : 1:50-1:500
Immunofluorescence (IF)/ICC	IF/ICC : 1:200-1:800
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

Published Applications

KD/KO	See 9 publications below
WB	See 21 publications below
IHC	See 10 publications below
IF	See 3 publications below

Product Information

12658-1-AP targets P4HA1 in WB, IHC, IF/ICC, IP, ELISA applications and shows reactivity with human, mouse, rat samples.

Tested Reactivity	human, mouse, rat
Cited Reactivity	human, mouse, rat
Host / Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	CatNo: Ag3292 Product name: Recombinant human P4HA1 protein Source: e coli.-derived, PGEX-4T Tag: GST Domain: 236-534 aa of BC034998 Sequence: PEHQRANGNLKYFEYIMAKEKDVNKSASDDQSDQKTTPKKKGVAVDYLPERQKYEMLCRGEGIKMTPRRQKKLFCRYHDGNRNPKFILAPAKQEDEWDKPRIIRFHDIISDAEIEIVKDLAKPRLRRATISNPITGDLETVHYRISKSAWLSGYENPVVSRINMRIQDLTGLDVSTAEELQVANYGVGGQYEPHFDFARKDEPDAFKELGTGNRIATWLFYMSDVSAGGATVFPEVGASVWPKKGTAVFWYNLFASGEGDYSTRHAACPVLVGNKWVSNKWLHERGQEFRRPCTLSELE 相同性解析による交差性が予測される生物種
Full Name	prolyl 4-hydroxylase, alpha polypeptide I
Calculated molecular weight	534 aa, 61 kDa
Observed molecular weight	61 kDa
GenBank accession number	BC034998
Gene Symbol	P4HA1
Gene ID (NCBI)	5033
RRID	AB_2283162
Conjugate	Unconjugated
Form
Form	Liquid
Purification Method	Antigen affinity purification
UNIPROT ID	P13674
Storage Buffer	PBS with 0.02% sodium azide and 50% glycerol{{ptg:BufferTemp}}7.3
Storage Conditions	Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

Background Information

P4HA1 (Prolyl 4-hydroxylase subunit alpha-1) is also known as P4HA and belongs to the P4HA family, which plays a central role in collagen synthesis. P4HA1 catalyzes the post-translational formation of 4-hydroxyproline in -Xaa-Pro-Gly- sequences in collagens and other proteins. The gene encodes a polypeptide of 517 amino acid residues and a signal peptide of 17 amino acids and the full length protein has two glycosylation sites (uniprot). It can exist as a heteromer, dimer or tetramer (refer to: GENATLAS). P4HA1 has some isoforms with the MW 61-63 kDa and 57-59 kDa.

Protocols

Product Specific Protocols
IF protocol for P4HA1 antibody 12658-1-AP	Download protocol
IHC protocol for P4HA1 antibody 12658-1-AP	Download protocol
IP protocol for P4HA1 antibody 12658-1-AP	Download protocol
WB protocol for P4HA1 antibody 12658-1-AP	Download protocol

Standard Protocols
Click here to view our Standard Protocols

Publications

Species	Application	Title
human	WB	Nature Cotranslational prolyl hydroxylation is essential for flavivirus biogenesis. Authors - Ranen Aviner View Article
rat,mouse	WB	Cell Death Differ Developmental growth plate cartilage formation suppressed by artificial light at night via inhibiting BMAL1-driven collagen hydroxylation Authors - Guangjin Chen View Article
mouse,human	WB	J Exp Med IL-17-induced HIF1α drives resistance to anti-PD-L1 via fibroblast-mediated immune exclusion. Authors - Xing Chen View Article
human	WB,IHC	Oncotarget The miR-124-prolyl hydroxylase P4HA1-MMP1 axis plays a critical role in prostate cancer progression. Authors - Balabhadrapatruni V S K Chakravarthi KD Validated View Article
human	WB,IHC,IF	Mol Oncol Prolyl 4-hydroxylase subunit alpha 1 (P4HA1) is a biomarker of poor prognosis in primary melanomas, and its depletion inhibits melanoma cell invasion and disrupts tumor blood vessel walls. Authors - Johanna Eriksson KD Validated View Article
human	WB	Aging (Albany NY) Identification of a novel glycolysis-related gene signature for predicting the prognosis of osteosarcoma patients. Authors - Mengkai Yang KD Validated View Article

Target

Host

Species Reactivity

Species Reactivity

Conjugate

Conjugate

Validation Data Gallery

WB analysis using 12658-1-AP

WB analysis of NIH/3T3 using 12658-1-AP

WB analysis of human heart using 12658-1-AP

WB analysis of rat skin using 12658-1-AP

WB analysis of HT-1080 using 12658-1-AP

IP experiment of A431 using 12658-1-AP

IHC staining of human pancreas cancer using 12658-1-AP

IHC staining of human pancreas cancer using 12658-1-AP

IHC staining of human breast cancer using 12658-1-AP

IHC staining of human breast cancer using 12658-1-AP

IF Staining of U2OS using 12658-1-AP

P4HA1 Polyclonal antibody

P4HA1 Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

-/ -

Tested Applications

Recommended dilution

Published Applications

Product Information

Background Information

Protocols

Publications

Cotranslational prolyl hydroxylation is essential for flavivirus biogenesis.

Developmental growth plate cartilage formation suppressed by artificial light at night via inhibiting BMAL1-driven collagen hydroxylation

IL-17-induced HIF1α drives resistance to anti-PD-L1 via fibroblast-mediated immune exclusion.

The miR-124-prolyl hydroxylase P4HA1-MMP1 axis plays a critical role in prostate cancer progression.

Prolyl 4-hydroxylase subunit alpha 1 (P4HA1) is a biomarker of poor prognosis in primary melanomas, and its depletion inhibits melanoma cell invasion and disrupts tumor blood vessel walls.

Identification of a novel glycolysis-related gene signature for predicting the prognosis of osteosarcoma patients.