WB
Western blot analysis using GRP78/Bip antibody against Huvec(1)Thp-1(2),Mcf-7(3),Hela(4),Hepg2(5),A549(6),Hek293(7),Jurkat(8),Cos-7(9),K562(10),A431(11) lysate . 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG (H&L) - HRP（OM644366）, at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60SIF-P
Immunohistochemical analysis of paraffin-embedded human Kidney tissues using GRP78/Bip antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:150 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:150 and incubate for one hour. Tyramine labeled with 488 (Green,OM642679) was used as chromogenic agent, and DAPI(Blue,OM642679) was used for double dyeing, and the anti-fluorescence attenuation tablets were sealed.IHC
Immunohistochemical analysis of paraffin-embedded breast cancer tissues using GRP78/Bip antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.IHC
Immunohistochemical analysis of paraffin-embedded Kidney tissues using GRP78/Bip antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.IHC
Immunohistochemical analysis of paraffin-embedded liver cancer tissues using GRP78/Bip antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.IHC
Immunohistochemical analysis of paraffin-embedded lung cancer tissues using GRP78/Bip antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.ICC/IF
Immunofluorescence analysis of C2C12 cells using GRP78/Bip antibody (green). Blue: DAPI fluorescent DNA dye. Red: Actin filaments have been labeled with Omnimabs® 594-Phalloidin.Cells are fixed in 4% paraformaldehyde at room temperature for 15 minutes. Then, they are permeabilized with a PBS (OM750003) solution containing 0.1% Triton X-100(OM750021) at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum(OM760028) at room temperature for 1 hour.The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, Omnimabs® 488 Goat Ant-Mouse IgG(H&L) (Green,OM643485), is diluted at a ratio of 1:400 and incubated with the cells for 1 hour.Nuclear DNA is labeled with DAPI (Blue,OM643160). F-actin is stained with Omnimabs® 594-Phalloidin (Red,OM750007) diluted 1:100 for 30 minutesELISA
Blue line: Antibody; Orange line: Isotype control antibody| Product Name | Anti-GRP78/Bip antibody,pbs only |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Mouse |
| Tested Applications | ELISAICC/IFIF-PIHCWB |
| WB:1:1000-1:2000 IF-P:1:200-1:2000 IHC:1:200-1:2000 ICC/IF:1:100-1:500 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Symbol | HSPA5 |
|---|---|
| Gene Synonyms | baffled BIP BiP/Grp78 D2Wsu141e D2Wsu17e GRP 78 GRP78 HEL-S-89n Hsce70 hspa5 hspa5a mBiP SEZ-7 Sez7 XAP-1 XELAEV_180384511mg YJL034W |
| Gene Full Name | heat shock protein family A (Hsp70) member 5 |
| Gene Summary | The protein encoded by this gene is a member of the heat shock protein 70 (HSP70) family. This protein localizes to the lumen of the endoplasmic reticulum (ER) where it operates as a typical HSP70 chaperone involved in the folding and assembly of proteins in the ER and is a master regulator of ER homeostasis. During cellular stress, as during viral infection or tumorogenesis, this protein interacts with the transmembrane stress sensor proteins PERK (protein kinase R-like endoplasmic reticulum kinase), IRE1 (inositol-requiring kinase 1), and ATF6 (activating transcription factor 6) where it acts as a repressor of the unfolded protein response (UPR) and also plays a role in cellular apoptosis and senescence. Elevated expression and atypical translocation of this protein to the cell surface has been reported in viral infections and some types of cancer cells. At the cell surface this protein may facilitate viral attachment and entry to host cells. This gene is a therapeutic target for the treatment of coronavirus diseases and chemoresistant cancers. [provided by RefSeq, Jul 2020] |
| Alternative Names | 78 kDa glucose-regulated protein binding-immunoglobulin protein BiP BiP protein chaperone endoplasmic reticulum lumenal Ca(2+)-binding protein grp78 epididymis secretory sperm binding protein Li 89n FLJ26106 glucose regulated protein 78 kDa glucose-regulated protein glucose-regulated protein (grp78) glucose-regulated protein 78 glucose-regulated protein 78kDa GRP-78 heat shock 70 kDa protein 5 heat shock 70 kDa protein 5a heat shock 70kD protein 5 heat shock 70kD protein 5 (glucose-regulated protein 78kD) heat shock 70kDa protein 5 (glucose-regulated protein) heat shock 70kDa protein 5 (glucose-regulated protein 78kDa) heat shock protein 5 heat shock protein 70 family protein 5 heat shock protein family A member 5 heavy-chain binding protein BiP HSP70 family protein 5 HSPA 5 immunoglobulin binding protein immunoglobulin heavy chain binding protein immunoglobulin heavy chain-binding protein preimmunoglobulin heavy chain binding protein steroidogenesis-activator polypeptide XAP-1 antigen |
| Molecular Weight(MW) | 73kDa |
| Cellular Localization | Endoplasmic reticulum lumen,Melanosome,Cytoplasm,Cell surface. |
| SwissProt ID | P11021 |
|---|
WB
Western blot analysis using GRP78/Bip antibody against Huvec(1)Thp-1(2),Mcf-7(3),Hela(4),Hepg2(5),A549(6),Hek293(7),Jurkat(8),Cos-7(9),K562(10),A431(11) lysate . 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG (H&L) - HRP（OM644366）, at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60S
IF-P
Immunohistochemical analysis of paraffin-embedded human Kidney tissues using GRP78/Bip antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:150 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:150 and incubate for one hour. Tyramine labeled with 488 (Green,OM642679) was used as chromogenic agent, and DAPI(Blue,OM642679) was used for double dyeing, and the anti-fluorescence attenuation tablets were sealed.
IHC
Immunohistochemical analysis of paraffin-embedded breast cancer tissues using GRP78/Bip antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
IHC
Immunohistochemical analysis of paraffin-embedded Kidney tissues using GRP78/Bip antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
IHC
Immunohistochemical analysis of paraffin-embedded liver cancer tissues using GRP78/Bip antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
IHC
Immunohistochemical analysis of paraffin-embedded lung cancer tissues using GRP78/Bip antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
ICC/IF
Immunofluorescence analysis of C2C12 cells using GRP78/Bip antibody (green). Blue: DAPI fluorescent DNA dye. Red: Actin filaments have been labeled with Omnimabs® 594-Phalloidin.Cells are fixed in 4% paraformaldehyde at room temperature for 15 minutes. Then, they are permeabilized with a PBS (OM750003) solution containing 0.1% Triton X-100(OM750021) at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum(OM760028) at room temperature for 1 hour.The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, Omnimabs® 488 Goat Ant-Mouse IgG(H&L) (Green,OM643485), is diluted at a ratio of 1:400 and incubated with the cells for 1 hour.Nuclear DNA is labeled with DAPI (Blue,OM643160). F-actin is stained with Omnimabs® 594-Phalloidin (Red,OM750007) diluted 1:100 for 30 minutes
ELISA
Blue line: Antibody; Orange line: Isotype control antibody| Application Notes | WB:1:1000-1:2000 IF-P:1:200-1:2000 IHC:1:200-1:2000 ICC/IF:1:100-1:500 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term. Avoid freeze / thaw cycle. |
| Storage Buffer | PBS ONLY |
