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GRP78 / BIP Mouse Monoclonal Antibody [C9-9]

Usd: 350

Specification

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Catalog# M1506-2

GRP78 / BIP Mouse Monoclonal Antibody [C9-9]

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Predicted reactivity

  • Zebrafish

Predicted species support after-sales service
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

GRP78 / BIP Mouse Monoclonal Antibody [C9-9]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Recombinant protein within human GRP78 aa 100-400.

Species Reactivity

Human, Mouse, Rat (Predicted: Zebrafish)

Predicted species support after-sales service

Validated Applications

WB, IF-Cell, IHC-P

Molecular Weight

Predicted band size: 72 kDa

Positive Control

MCF7 cell lysate, HepG2 cell lysate, Human brain tissue lysate, Mouse brain tissue 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, human liver tissue, mouse liver tissue, rat liver tissue, HepG2.

Conjugation

unconjugated

Clone Number

C9-9

RRID

AB_3073113

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:5,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:1,000

Target

Function

Binding immunoglobulin protein (BiP) also known as 78 kDa glucose-regulated protein (GRP-78) or heat shock 70 kDa protein 5 (HSPA5) is a protein that in humans is encoded by the HSPA5 gene. BiP is a HSP70 molecular chaperone located in the lumen of the endoplasmic reticulum (ER) that binds newly synthesized proteins as they are translocated into the ER, and maintains them in a state competent for subsequent folding and oligomerization. BiP is also an essential component of the translocation machinery, as well as playing a role in retrograde transport across the ER membrane of aberrant proteins destined for degradation by the proteasome. Like many stress and heat shock proteins, BiP/GRP78 has potent immunological activity when released from the internal environment of the cell into the extracelluar space.specifically, it feeds anti-inflammatory and pro-resolutory signals into immune networks, thus helping to resolve inflammation.

Background References

1. "Human XTP3-B forms an endoplasmic reticulum quality control scaffold with the HRD1-SEL1L ubiquitin ligase complex and BiP." Hosokawa N., Wada I., Nagasawa K., Moriyama T., Okawa K., Nagata K. J. Biol. Chem. 283:20914-20924(2008)

2. "Crystal structures of the ATPase domains of four human Hsp70 isoforms: HSPA1L/Hsp70-hom, HSPA2/Hsp70-2, HSPA6/Hsp70B\', and HSPA5/BiP/GRP78." Wisniewska M., Karlberg T., Lehtio L., Johansson I., Kotenyova T., Moche M., Schuler H. PLoS ONE 5:E8625-E8625(2010)

3. "Adenosine-derived inhibitors of 78 kDa glucose regulated protein (Grp78) ATPase: insights into isoform selectivity." Macias A.T., Williamson D.S., Allen N., Borgognoni J., Clay A., Daniels Z., Dokurno P., Drysdale M.J., Francis G.L., Graham C.J., Howes R., Matassova N., Murray J.B., Parsons R., Shaw T., Surgenor A.E., Terry L., Wang Y., Wood M., Massey A.J. J. Med. Chem. 54:4034-4041(2011)

Sequence Similarity

Belongs to the heat shock protein 70 family.

Post-translational Modification

AMPylated by FICD. In unstressed cells, AMPylation at Thr-518 by FICD inactivates the chaperome activity: AMPylated form is locked in a relatively inert state and only weakly stimulated by J domain-containing proteins (By similarity). In response to endoplasmic reticulum stress, de-AMPylation by the same protein, FICD, restores the chaperone activity (By similarity).

Subcellular Location

Cytoplasm, endoplasmic reticulum lumen

UNIPROT

SwissProt: P11021 Human

SwissProt: P20029 Mouse

SwissProt: P06761 Rat

Synonyms

78 kDa glucose regulated protein antibody

78 kDa glucose-regulated protein antibody

AL022860 antibody

AU019543 antibody

BIP antibody

D2Wsu141e antibody

D2Wsu17e antibody

Endoplasmic reticulum lumenal Ca(2+)-binding protein grp78 antibody

Endoplasmic reticulum lumenal Ca2+ binding protein grp78 antibody

Epididymis secretory sperm binding protein Li 89n antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of GRP78 / BIP on different lysates with Mouse anti-GRP78 / BIP antibody (<a href="/products/M1506-2" style="font-weight: bold;text-decoration: underline;">M1506-2</a>) at 1/1,000 dilution.<br /><br />Lane 1: MCF7 cell lysate (15 µg/Lane)<br />Lane 2: HepG2 cell lysate (15 µg/Lane)<br />Lane 3: Human brain tissue lysate (30 µg/Lane)<br />Lane 4: Mouse brain tissue lysate (30 µg/Lane)<br />Lane 5: U-87 MG cell lysate (30 µg/Lane)<br />Lane 6: RAW264.7 cell lysate (30 µg/Lane)<br />Lane 7: RAW264.7 treated with 300nM Thapsigargin for 18 hours cell lysate (30 µg/Lane)<br />Lane 8: Mouse liver tissue lysate (30 µg/Lane)<br />Lane 9: Rat liver tissue lysate (30 µg/Lane)<br />Lane 10: Rat pancreas tissue lysate (30 µg/Lane)<br /><br />Predicted band size: 72 kDa<br />Observed band size: 70/72 kDa<br /><br />Exposure time: Lane 1-4: 11 seconds; Lane 5-10: 4 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/M1506-2" style="font-weight: bold;text-decoration: underline;">M1506-2</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of GRP78 / BIP on different lysates with Mouse anti-GRP78 / BIP antibody (M1506-2) at 1/1,000 dilution.

    Lane 1: MCF7 cell lysate (15 µg/Lane)
    Lane 2: HepG2 cell lysate (15 µg/Lane)
    Lane 3: Human brain tissue lysate (30 µg/Lane)
    Lane 4: Mouse brain tissue lysate (30 µg/Lane)
    Lane 5: U-87 MG cell lysate (30 µg/Lane)
    Lane 6: RAW264.7 cell lysate (30 µg/Lane)
    Lane 7: RAW264.7 treated with 300nM Thapsigargin for 18 hours cell lysate (30 µg/Lane)
    Lane 8: Mouse liver tissue lysate (30 µg/Lane)
    Lane 9: Rat liver tissue lysate (30 µg/Lane)
    Lane 10: Rat pancreas tissue lysate (30 µg/Lane)

    Predicted band size: 72 kDa
    Observed band size: 70/72 kDa

    Exposure time: Lane 1-4: 11 seconds; Lane 5-10: 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 (M1506-2) 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.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Mouse anti-GRP78 / BIP antibody (<a href="/products/M1506-2" style="font-weight: bold;text-decoration: underline;">M1506-2</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/M1506-2" style="font-weight: bold;text-decoration: underline;">M1506-2</a>) 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.

    Immunohistochemical analysis of paraffin-embedded human liver tissue with Mouse anti-GRP78 / BIP antibody (M1506-2) 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 (M1506-2) 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.

  • Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Mouse anti-GRP78 / BIP antibody (<a href="/products/M1506-2" style="font-weight: bold;text-decoration: underline;">M1506-2</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/M1506-2" style="font-weight: bold;text-decoration: underline;">M1506-2</a>) 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.

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Mouse anti-GRP78 / BIP antibody (M1506-2) 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 (M1506-2) 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.

  • Immunohistochemical analysis of paraffin-embedded rat liver tissue with Mouse anti-GRP78 / BIP antibody (<a href="/products/M1506-2" style="font-weight: bold;text-decoration: underline;">M1506-2</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/M1506-2" style="font-weight: bold;text-decoration: underline;">M1506-2</a>) 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.

    Immunohistochemical analysis of paraffin-embedded rat liver tissue with Mouse anti-GRP78 / BIP antibody (M1506-2) 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 (M1506-2) 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.

  • Immunocytochemistry analysis of HepG2 cells labeling GRP78 / BIP with Mouse anti-GRP78 / BIP antibody (<a href="/products/M1506-2" style="font-weight: bold;text-decoration: underline;">M1506-2</a>) at 1/100 dilution.<br /><br />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 (<a href="/products/M1506-2" style="font-weight: bold;text-decoration: underline;">M1506-2</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) 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.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HepG2 cells labeling GRP78 / BIP with Mouse anti-GRP78 / BIP antibody (M1506-2) at 1/100 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 (M1506-2) at 1/100 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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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