Anti-P4HA1 Antibody, clone 1C14 ZooMAb^® Rabbit Monoclonal

SIGMA/ZRB1744 - recombinant, expressed in HEK 293 cells

Synonym: Prolyl 4-hydroxylase subunit alpha-1;EC:1.14.11.2;4-PH alpha-1;Procollagen-proline;2-oxoglutarate-4-dioxygenase subunit alpha-1

Product Type: Chemical

Catalog Number	PKG Qty.	Price	Quantity


45-ZRB1744-25UL		No Price
45-ZRB1744-4X25UL		No Price

Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Placenta (A and B) and Negative control (no primary) (C and D) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1744, Anti-P4HA1, clone 1C14 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in mesenchyme of chorionic villi of Human Placenta sections.

Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of U2OS cells was performed using a 1:1,000 dilution of Cat. No. ZRB1744, Anti-P4HA1, clone 1C14 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the endoplasmic reticulum.

Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from HEK293 (Lane 1) and U2OS (Lane 2) cells were probed with Cat. No. ZRB1744, Anti-P4HA1, clone 1C14 ZooMAb® Rabbit Monoclonal (1:10,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates P4HA1 (~59 kDa).

Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or P4HA1 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1744, Anti-P4HA1, clone 1C14 ZooMAb® Rabbit Monoclonal. The binding of this antibody to P4HA1 peptide displayed a KD of 9.1 x 10-8, whereas binding to non-specific control peptide displayed a KD of 2.0 x 10-3.

12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.

12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.

12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.

Properties
Descriptions
Basic Data

antibody form	purified antibody
antibody product type	primary antibodies
biological source	rabbit
clone	1C14, recombinant monoclonal
conjugate	unconjugated
description	recombinant, expressed in HEK 293 cells
enhanced validation	recombinant expression Learn more about Antibody Enhanced Validation
epitope sequence	Internal
form	lyophilized
greener alternative category	Aligned
greener alternative product characteristics	Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry .
isotype	IgG
mol wt	calculated mol wt 61.05 kDa
	observed mol wt ~59 kDa
packaging	antibody small pack of 25 μL
product line	ZooMAb^® learn more
Protein ID accession no.	NP_001017962
purified by	using Protein A
Quality Level	200
recombinant	expressed in HEK 293 cells
shipped in	ambient
species reactivity	human
species reactivity (predicted by homology)	monkey
storage temp.	2-8°C
technique(s)	affinity binding assay: suitable
	immunocytochemistry: suitable
	immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable
	western blot: suitable
UniProt accession no.	P13674

Application:	Quality Control Testing Evaluated by Western Blotting in HEK293 cell lysate. Western Blotting Analysis: A 1:10,000 dilution of this antibody detected P4HA1 in HEK293 cell lysate. Tested applications Western Blotting Analysis: A 1:10,000 dilution from a representative lot detected P4HA1 in U2OS cell lysate. Immunocytochemistry Analysis: A 1:1,000 dilution from a representative lot detected P4HA1 in U2OS cells. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected P4HA1 in human placenta tissue sections. Affinity Binding Assay: A representative lot of this antibody bound P4HA1 with a KD of 9.1 x 10-8 in an affinity binding assay. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user
Disclaimer:	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
General description:	We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information.
General description:	ZooMAb^® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb^® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb^® antibodies are reliably available and ready to ship when you need them.
Immunogen:	KLH-conjugated linear peptide corresponding to 25 amino acids from the internal region of human Prolyl 4-hydroxylase subunit alpha-1 (P4HA1)
Legal Information:	ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany
Physical form:	Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL.
Reconstitution:	300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Specificity:	Clone 1C14 is a ZooMAb^® Rabbit recombinant monoclonal antibody that specifically detects Prolyl 4-hydroxylase subunit alpha-1 (P4HA1), It targets an epitope within 25 amino acids from the internal region.
Storage and Stability:	Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.
Target description:	Prolyl 4-hydroxylase subunit alpha-1 (UniProt: P13674; also known as EC:1.14.11.2, 4-PH alpha-1, Procollagen-proline,2-oxoglutarate-4-dioxygenase subunit alpha-1, P4HA1) is encoded by the P4HA1 (also known as P4HA) gene (Gene ID: 5033) in human. Prolyl 4-hydroxylase is a key enzyme in collagen synthesis and is composed of two identical a subunits and two b subunits. In collagen and related proteins, prolyl 4-hydroxylase is reported to catalyze the formation of 4-hydroxyproline, which is essential to the proper three-dimensional folding of newly synthesized procollagen chains. Three P4HA isoforms (P4HA1-3) have been identified in mammalian cells and P4HA1 is the major isoform described in most cell types and tissues and contributes to the majority of the prolyl 4-hydroxylase activity. It is localized in endoplasmic reticulum lumen and catalyzes the post-translational formation of 4-hydroxyproline in -Xaa-Pro-Gly- sequences in collagens and other proteins. Depletion of P4HA1 in melanoma cells is reported to reduce cell adhesion, invasion, and viability in vitro. In melanoma xenograft models, P4HA1 knockdown reduces melanoma tumor invasion and the deposition of collagens, particularly type IV collagen, in the interstitial extracellular matrix and in the basement membranes of tumor blood vessels. This can lead to vessel wall rupture and hemorrhages. Significantly higher levels of P4HA1 are reported in subjects with lung adenocarcinomas and relate to worse overall survival. This ZooMAb^® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Zhou, H., et al. (2020). Biomed. Res. Int. 2020, Article ID 8024138; Eriksson, J., et al. (2020). Mol. Oncol. 14(4); 742-762).

Storage Temp.	2-8°C
UNSPSC	12352203

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