M1505-13

GRP78 / BIP Mouse Monoclonal Antibody [2-7]

cat.: M1505-13

Product Type:	Mouse monoclonal IgG1, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, IF-Tissue, FC
Clonality:	Monoclonal
Clone number:	2-7

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, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 78 kDa
Isotype:	IgG1
Immunogen:	Synthetic peptide within Human GRP78 aa 605-654 / 654.
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, hybrid fish (crucian-carp) heart tissue lysates.
Subcellular location:	Cytoplasm, endoplasmic reticulum lumen
Recommended Dilutions: WB IF-Cell IF-Tissue FC	1:1,000-1:5,000 1:200 1:50-1:200 1;1,000
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

Fig1: Western blot analysis of GRP78 / BIP on different lysates with Mouse anti-GRP78 / BIP antibody (M1505-13) 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: 4 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 (M1505-13) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of GRP78 / BIP on hybrid fish (crucian-carp) heart tissue lysate using anti-GRP78 / BIP antibody at 1/500 dilution.

	Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Mouse anti-GRP78 / BIP antibody (M1505-13) at 1/1,000 dilution. 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 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1505-13) at 1/1,000 dilution for 1 hour 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human liver tissue with Mouse anti-GRP78 / BIP antibody (M1505-13) at 1/1,000 dilution. 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 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1505-13) at 1/1,000 dilution for 1 hour 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Mouse anti-GRP78 / BIP antibody (M1505-13) at 1/1,000 dilution. 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 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1505-13) at 1/1,000 dilution for 1 hour 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.

	Fig6: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Mouse anti-GRP78 / BIP antibody (M1505-13) at 1/1,000 dilution. 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 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1505-13) at 1/1,000 dilution for 1 hour 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.
	Fig7: Immunocytochemistry analysis of L-929 cells labeling GRP78 / BIP with Mouse anti-GRP78 / BIP antibody (M1505-13) at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-GRP78 / BIP antibody (M1505-13) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.
	Fig8: Immunocytochemistry analysis of RAW264.7 cells labeling GRP78 / BIP with Mouse anti-GRP78 / BIP antibody (M1505-13) at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-GRP78 / BIP antibody (M1505-13) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

Fig9: Immunocytochemistry analysis of PC-12 cells labeling GRP78 / BIP with Mouse anti-GRP78 / BIP antibody (M1505-13) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-GRP78 / BIP antibody (M1505-13) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.