ATP5A1 Recombinant antibody

ATP5A1 Recombinant Antibody for WB, IF, FC, IHC, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

Human, mouse, rat

Applications

WB, IF, FC, IHC, ELISA

Conjugate

Unconjugated

CloneNo.

4O12

Cat no : 82288-1-RR

Synonyms

ATP5A, ATP5A1, ATP5AL2, ATPM, hATP1, MOM2, OMR, ORM

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Validation Data Gallery

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  WB analysis using 82288-1-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 82288-1-RR (ATP5A1 antibody) at dilution of 1:30000 incubated at room temperature for 1.5 hours.

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  IHC staining of mouse brain using 82288-1-RR

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 82288-1-RR (ATP5A1 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse brain using 82288-1-RR

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 82288-1-RR (ATP5A1 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human liver using 82288-1-RR

  Immunohistochemical analysis of paraffin-embedded human liver tissue slide using 82288-1-RR (ATP5A1 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human liver using 82288-1-RR

  Immunohistochemical analysis of paraffin-embedded human liver tissue slide using 82288-1-RR (ATP5A1 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 82288-1-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed HeLa cells using ATP5A1 antibody (82288-1-RR, Clone: 4O12 ) at dilution of 1:1000 and CoraLite®594-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

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  IF Staining of HepG2 using 82288-1-RR

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using ATP5A1 antibody (82288-1-RR, Clone: 4O12 ) at dilution of 1:2000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

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  IF Staining of HeLa using 82288-1-RR

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using ATP5A1 antibody (82288-1-RR, Clone: 4O12 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

• 

  IF Staining of HeLa using 82288-1-RR

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using ATP5A1 antibody (82288-1-RR, Clone: 4O12 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

• 

  IF Staining of HeLa using 82288-1-RR

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using ATP5A1 antibody (82288-1-RR, Clone: 4O12 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  FC experiment of HeLa using 82288-1-RR

  1X10^6 HeLa cells were intracellularly stained with 0.4 ug Anti-Human ATP5A1 (82288-1-RR, Clone:4O12) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Isotype Control. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: HEK-293 cells, HeLa cells, HepG2 cells, MCF-7 cells, Jurkat cells, NIH/3T3 cells, HSC-T6 cells
• Positive IHC detected in: mouse brain tissue, human liver 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 detected in: HepG2 cells, HeLa cells
• Positive FC detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF): IF : 1:1100-1:4400
• Flow Cytometry (FC): FC : 0.40 ug per 10^6 cells in a 100 µl suspension

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.

Product Information

82288-1-RR targets ATP5A1 in WB, IF, FC, IHC, ELISA applications and shows reactivity with Human, mouse, rat samples.

• Tested Reactivity: Human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: ATP5A1 fusion protein Ag6385
• Full Name: ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit 1, cardiac muscle
• Calculated Molecular Weight: 60 kDa
• Observed Molecular Weight: 50-55 kDa
• GenBank Accession Number: BC064562
• Gene Symbol: ATP5A1
• Gene ID (NCBI): 498
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

The ATP5A1 gene encodes the α subunit of mitochondrial ATP synthase which produces ATP from ADP in the presence of a proton gradient across the membrane. The mitochondrial ATP synthase, also known as Complex V or F1F0 ATP synthase, is a multi-subunit enzyme complex consisting of two functional domains, the F1-containing the catalytic core and the Fo-containing the membrane proton channel. F0 domain has 10 subunits: a,b, c, d, e, f, g, OSCP, A6L, and F6. F1 is composed of subunits α, β, γ, δ, ε, and a loosely attached inhibitor protein IF1. Recently defect in ATP5A1 has been linked to the fatal neonatal mitochondrial encephalopathy. ATP5A1 is localized in the mitochondria and anti-ATP5A1 can be used as the loading control for mitochondrial or Complex V proteins. This antibody recognizes the endogenous ATP5A1 protein in lysates from various cell lines and tissues. The predicted MW of ATP5A1 is 60 kDa, while it undergoes the transit peptide cleavage to become a mature form around 50-55 kDa. Several isoforms of ATP5A1 exist due to the alternative splicing.