GRP75 Polyclonal antibody

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  WB analysis using 14887-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 14887-1-AP (GRP75 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

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  WB analysis of A549 cells using 14887-1-AP

  A549 cells (shcontrol and shRNA of GRP75)were subjected to SDS PAGE followed by western blot with 14887-1-AP (GRP75 antibody) at dilution of 1:500.

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  WB analysis of mouse testis using 14887-1-AP

  mouse testis tissue were subjected to SDS PAGE followed by western blot with 14887-1-AP (GRP75 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  IP experiment of mouse brain using 14887-1-AP

  IP result of anti-GRP75 (IP:14887-1-AP, 4ug; Detection:14887-1-AP 1:2000) with mouse brain tissue lysate 3680ug.

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  IP experiment of NIH/3T3 using 14887-1-AP

  IP result of anti-GRP75 (IP:14887-1-AP, 4ug; Detection:14887-1-AP 1:700) with NIH/3T3 cells lysate 1200ug.

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  IHC staining of human normal colon using 14887-1-AP

  Immunohistochemical analysis of paraffin-embedded human normal colon slide using 14887-1-AP (GRP75 antibody) at dilution of 1:4000 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HepG2 using 14887-1-AP

  Immunofluorescent analysis of HepG2 cells using 14887-1-AP (GRP75 antibody) at dilution of 1:25 and Rhodamine-Goat anti-Rabbit IgG.

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  IF Staining of HepG2 using 14887-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed HepG2 cells using GRP75 antibody (14887-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

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  FC experiment of HepG2 using 14887-1-AP

  1x10^6 HepG2 cells were intracellularly stained with 0.2 ug GRP75 Polyclonal antibody (14887-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.2 ug Rabbit IgG control Rabbit PolyAb (30000-0-AP) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

• Featured Product
• KD/KO Validated

GRP75 Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Cat No. 14887-1-AP

Publications(49)

Reviews (4)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat, zebrafish and More (1)

Applications

WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA

Synonyms

HSPA9, Heat shock protein family A member 9, Heat shock 70 kDa protein 9, GRP-75, CSA

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Tested Applications

• Positive WB detected in: HT-29 cells, A549 cells, mouse testis tissue, Jurkat cells, NIH/3T3 cells, mouse liver
• Positive IP detected in: mouse brain tissue, NIH/3T3 cells
• Positive IHC detected in: human normal colon; 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: HepG2 cells
• Positive FC (Intra) detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:2000-1:8000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.20 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.

Published Applications

• KD/KO: See 3 publications below
• WB: See 39 publications below
• IHC: See 4 publications below
• IF: See 11 publications below
• IP: See 3 publications below
• CoIP: See 1 publications below

Product Information

14887-1-AP targets GRP75 in WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA applications and shows reactivity with human, mouse, rat, zebrafish samples.

• Tested Reactivity: human, mouse, rat, zebrafish
• Cited Reactivity: human, mouse, rat, chicken, zebrafish
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: GRP75 fusion protein Ag6673
• Full Name: heat shock 70kDa protein 9 (mortalin)
• Calculated Molecular Weight: 74 kDa
• Observed Molecular Weight: 74-75 kDa
• GenBank Accession Number: BC000478
• Gene Symbol: GRP75
• Gene ID (NCBI): 3313
• RRID: AB_2120458
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• UNIPROT ID: P38646
• 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

GRP75 (also known as mortalin, HSPA9 or mt-Hsp70) is a constitutively expressed member of the HSPA (HSP70) family of heat-shock proteins. It is located in the mitochondrial matrix and anti-GRP75 is commonly used as the marker for mitochondria. It has been reported that GRP75 is enriched in cancer cells and contributes to carcinogenesis. In addition, decreased expression level of GRP75 has been found in neurodegenerative disorders like Parkinson's disease. This antibody recognized the endogenous GRP75 protein in NIH/3T3 cells. (PMID: 21640711, 22920904)

Publications

Species	Application	Title
human	WB	Mol Cell; The mitochondrial DNAJC co-chaperone TCAIM reduces α-ketoglutarate dehydrogenase protein levels to regulate metabolism; Authors - Wang Jiahui ; KD Validated
human	WB	Cell Metab; DNAJC19, a mitochondrial cochaperone associated with cardiomyopathy, forms a complex with prohibitins to regulate cardiolipin remodeling.; Authors - Ricarda Richter-Dennerlein
human	WB	Nat Commun; LONP1 and mtHSP70 cooperate to promote mitochondrial protein folding.; Authors - Chun-Shik Shin
mouse		EMBO Mol Med; Excess hydrogen sulfide and polysulfides production underlies a schizophrenia pathophysiology.; Authors - Masayuki Ide
mouse	WB	J Cell Biol; Loss of OMA1 delays neurodegeneration by preventing stress-induced OPA1 processing in mitochondria.; Authors - Anne Korwitz
rat	WB	Cell Death Dis; Pathological convergence of APP and SNCA deficiency in hippocampal degeneration of young rats; Authors - Yajie Wang

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Anastasia (Verified Customer) (08-07-2024)

Deyolked zebrafish embryos

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: Zebrafish embryos

GRP75 Antibody Western Blot validation (1:1000 dilution) in Zebrafish embryos (Cat no:14887-1-AP)

FH

Christina (Verified Customer) (11-14-2023)

I used this antibody for HeLa extracts in Western Blot and worked perfectly.

• Applications: Western Blot
• Primary Antibody Dilution: 1:5000
• Cell Tissue Type: HeLa cells

FH

Ning (Verified Customer) (02-27-2023)

Specifically probe endogenous GRP75 with a right MW.

• Applications: Western Blot
• Primary Antibody Dilution: 1:2000
• Cell Tissue Type: mouse brain lysate

GRP75 Antibody Western Blot validation (1:2000 dilution) in mouse brain lysate (Cat no:14887-1-AP)

FH

Staci (Verified Customer) (02-02-2023)

Antibody works very well with GRP75. Very little non-specific binding on immunoblot.

• Applications: Western Blot
• Primary Antibody Dilution: 1:1,000
• Cell Tissue Type: Mouse Islet protein lysates