GRP78/BIP Monoclonal antibody

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  WB analysis using 66574-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66574-1-Ig (GRP78/BIP antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated Alpha Tubulin Monoclonal antibody (HRP-66031) as loading control.

• 

  WB analysis using 66574-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66574-1-Ig (GRP78/BIP antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 66574-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66574-1-Ig (GRP78/BIP antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 66574-1-Ig

  Various cells were subjected to SDS PAGE followed by western blot with 66574-1-Ig (GRP78,BIP antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  IP experiment of SGC-7901 using 66574-1-Ig

  IP result of anti-GRP78/BIP (IP:66574-1-Ig, 5ug; Detection:66574-1-Ig 1:3000) with SGC-7901 cells lysate 2200 ug.

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  IHC staining of human liver cancer using 66574-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66574-1-Ig (GRP78,BIP antibody) at dilution of 1:1000 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human liver cancer using 66574-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66574-1-Ig (GRP78,BIP antibody) at dilution of 1:1000 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 66574-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66574-1-Ig (GRP78,BIP antibody) at dilution of 1:1000 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 66574-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66574-1-Ig (GRP78,BIP antibody) at dilution of 1:1000 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human colon cancer using 66574-1-Ig

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 66574-1-Ig (GRP78,BIP antibody) at dilution of 1:1000 (under 10x lens).

• 

  IHC staining of human colon cancer using 66574-1-Ig

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 66574-1-Ig (GRP78,BIP antibody) at dilution of 1:1000 (under 40x lens).

• 

  IF Staining of U2OS using 66574-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed thapsigargin treated U2OS cells using GRP78/BIP antibody (66574-1-Ig, Clone: 1D6F7 ) at dilution of 1:800 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) (SA00013-1), CL594-phalloidin (red).

• Featured Product
• KD/KO Validated

GRP78/BIP Monoclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Cat No. 66574-1-Ig

Clone No.1D6F7

Publications(94)

Reviews (1)

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat and More (3)

Applications

WB, IHC, IF/ICC, IP, CoIP, ELISA

Synonyms

BIP, GRP78, HSPA5, 1D6F7, 78 kDa glucose-regulated protein

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Tested Applications

• Positive WB detected in: HepG2 cells, HeLa cells, A549 cells, MCF-7 cells, Jurkat cells, HSCT6 cells, HSC-T6 cells, PC-12 cells, NIH/3T3 cells, SP2/0 cell, 4T1 cells
• Positive IP detected in: SGC-7901 cells
• Positive IHC detected in: human liver cancer tissue, human colon 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: thapsigargin treated U2OS 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 : 1:500-1:2000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:400-1:1600

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 1 publications below
• WB: See 81 publications below
• IHC: See 10 publications below
• IF: See 22 publications below
• IP: See 3 publications below
• CoIP: See 3 publications below

Product Information

66574-1-Ig targets GRP78/BIP in WB, IHC, IF/ICC, IP, CoIP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, pig, chicken, zebrafish
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: GRP78/BIP fusion protein Ag2188
• Full Name: heat shock 70kDa protein 5 (glucose-regulated protein, 78kDa)
• Calculated Molecular Weight: 78 kDa
• Observed Molecular Weight: 70-78 kDa
• GenBank Accession Number: BC020235
• Gene Symbol: GRP78/BIP
• Gene ID (NCBI): 3309
• RRID: AB_2881934
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• UNIPROT ID: P11021
• 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

GRP78 (HSPA5), also referred to as 'immunoglobulin heavy chain-binding protein' (BiP), is a member of the heat-shock protein-70 (HSP70) family and is involved in the folding and assembly of proteins in the endoplasmic reticulum (ER).It is a constitutively expressed resident protein of the ER in all eukaryotic cells. Recently it has been reported that GRP78 is associated with apoptosis or inhibition of cancer cell growth.

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
mouse	WB	J Clin Invest; Sec13 promotes oligodendrocyte differentiation and myelin repair through autocrine pleiotrophin signaling.; Authors - Zhixiong Liu
human	WB	Redox Biol; Mecheliolide elicits ROS-mediated ERS driven immunogenic cell death in hepatocellular carcinoma.; Authors - Zhongren Xu
	WB	Food Chem; Crosstalk between mitochondrial dysfunction and endoplasmic reticulum stress in apoptosis: Implications for grass carp flesh quality during cold storage; Authors - Xin Xiong
mouse,human	WB,IHC,IF	Int J Biol Macromol; p20BAP31 promotes cell apoptosis via interaction with GRP78 and activating the PERK pathway in colorectal cancer; Authors - Xiaohan Jiang
mouse,human	WB	Phytomedicine; Asperosaponin VI protects alcohol-induced hepatic steatosis and injury via regulating lipid metabolism and ER stress; Authors - Linlin Wei
	WB	J Cell Biol; LET-767 determines lipid droplet protein targeting and lipid homeostasis; Authors - Lin Fu

Reviews

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

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FH

Udesh (Verified Customer) (08-19-2024)

Worked well at 1:2000, 5ug lysate

• Applications: Western Blot
• Primary Antibody Dilution: 1:2000
• Cell Tissue Type: MSCs

GRP78/BIP Antibody Western Blot validation (1:2000 dilution) in MSCs (Cat no:66574-1-Ig)