GRP78 / BIP Rabbit Polyclonal Antibody
cat.: ER1706-50
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Polyclonal
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Immunogen affinity purified.
Molecular weight: Predicted band size: 72 kDa
Isotype: IgG
Immunogen: Synthetic peptide within N-terminal human GRP78.
Positive control: L-929 cell lysate, U-87 MG cell lysate, RAW264.7 cell lysate, RAW264.7 treated with 300nM Thapsigargin for 18 hours cell lysate, mouse liver tissue lysate, rat liver tissue lysate, rat pancreas tissue lysate, Hela, SH-SY5Y, A431, HepG2, HUVEC, rat brain tissue, human liver tissue, mouse cerebellum tissue, human placenta tissue, human stomach carcinoma tissue, Jurkat.
Subcellular location: Endoplasmic reticulum. Cytoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:1,000-1:5,000
1:50-1:200
1:50-1:600
1:50-1:100
Uniprot #: SwissProt: P11021 Human | P20029 Mouse | P06761 Rat
Alternative names: 78 kDa glucose regulated protein 78 kDa glucose-regulated protein AL022860 AU019543 BIP D2Wsu141e D2Wsu17e Endoplasmic reticulum lumenal Ca(2+)-binding protein grp78 Endoplasmic reticulum lumenal Ca2+ binding protein grp78 Epididymis secretory sperm binding protein Li 89n FLJ26106 Glucose Regulated Protein 78kDa GRP 78 GRP-78 GRP78 GRP78_HUMAN Heat shock 70 kDa protein 5 Heat Shock 70kDa Protein 5 Heat shock protein family A (Hsp70) member 5 HEL S 89n Hsce70 HSPA 5 HSPA5 Immunoglobulin Heavy Chain Binding Protein Immunoglobulin heavy chain-binding protein mBiP MIF2 Sez7
Images
ER1706-50_1.jpg Fig1: Western blot analysis of GRP78 / BIP on different lysates with Rabbit anti-GRP78 / BIP antibody (ER1706-50) at 1/1,000 dilution.

Lane 1: L-929 cell lysate
Lane 2: U-87 MG cell lysate
Lane 3: RAW264.7 cell lysate
Lane 4: RAW264.7 treated with 300nM Thapsigargin for 18 hours cell lysate
Lane 5: Mouse liver tissue lysate
Lane 6: Rat liver tissue lysate
Lane 7: Rat pancreas tissue lysate

Lysates/proteins at 30 µg/Lane.

Predicted band size: 72 kDa
Observed band size: 72 kDa

Exposure time: 6 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ER1706-50) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ER1706-50_2.jpg Fig2: ICC staining of GRP78 / BIP in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ER1706-50, 1/200) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ER1706-50_3.jpg Fig3: ICC staining of GRP78 / BIP in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ER1706-50, 1/200) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ER1706-50_4.jpg Fig4: ICC staining of GRP78 / BIP in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ER1706-50, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ER1706-50_5.jpg Fig5: ICC staining of GRP78 / BIP in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ER1706-50, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ER1706-50_6.jpg Fig6: ICC staining of GRP78 / BIP in HUVEC cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ER1706-50, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ER1706-50_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-GRP78 / BIP antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1706-50, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ER1706-50_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-GRP78 / BIP antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1706-50, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ER1706-50_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue using anti-GRP78 / BIP antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1706-50, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ER1706-50_10.png Fig10: Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-GRP78 / BIP antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1706-50, 1/600) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ER1706-50_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue using anti-GRP78 / BIP antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1706-50, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ER1706-50_12.jpg Fig12: Flow cytometric analysis of GRP78 / BIP was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (ER1706-50, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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