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Tbp7 Recombinant Rabbit Monoclonal Antibody [JE62-72]

Usd: 385

Specification

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Catalog# HA721435

Tbp7 Recombinant Rabbit Monoclonal Antibody [JE62-72]

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

Tbp7 Recombinant Rabbit Monoclonal Antibody [JE62-72]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Tbp7 aa 369-418.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Cell, FC

Molecular Weight

Predicted band size: 47 kDa

Positive Control

K-562 cell lysate, HEK-293 cell lysate, HeLa cell lysate, U-2 OS cell lysate, HepG2 cell lysate, MCF7 cell lysate, Jurkat cell lysate, mouse testis tissue lysate, rat testis tissue lysate, human colon carcinoma tissue, mouse cerebellum tissue, rat cerebellum tissue, U-2 OS.

Conjugation

unconjugated

Clone Number

JE62-72

RRID

AB_3072551

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

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IHC-P

  • 1:1,000-1:2,000

  • IF-Cell

  • 1:100

  • FC

  • 1:500-1:1,000

Target

Function

26S protease regulatory subunit 6B, also known as 26S proteasome AAA-ATPase subunit Rpt3, is an enzyme that in humans is encoded by the PSMC4 gene. This protein is one of the 19 essential subunits of a complete assembled 19S proteasome complex Six 26S proteasome AAA-ATPase subunits (Rpt1, Rpt2, Rpt3 (this protein), Rpt4, Rpt5, and Rpt6) together with four non-ATPase subunits (Rpn1, Rpn2, Rpn10, and Rpn13) form the base sub complex of 19S regulatory particle for proteasome complex. The gene PSMC4 encodes one of the ATPase subunits, a member of the triple-A family of ATPases which have a chaperone-like activity. This subunit has been shown to interact with an orphan member of the nuclear hormone receptor superfamily highly expressed in liver, and with gankyrin, a liver oncoprotein. Two transcript variants encoding different isoforms have been identified. The human PSMC3 gene has 11 exons and locates at chromosome band 19q13.11-q13.13. The human protein 26S protease regulatory subunit 6B is 47kDa in size and composed of 418 amino acids. The calculated theoretical pI of this protein is 5.09.

Background References

1. Kao TJ et al. Prognoses and genomic analyses of proteasome 26S subunit, ATPase (PSMC) family genes in clinical breast cancer. Aging (Albany NY). 2021 Jul

2. Ullah MA et al. Evaluating the Prognostic and Therapeutic Potentials of the Proteasome 26S Subunit, ATPase (PSMC) Family of Genes in Lung Adenocarcinoma: A Database Mining Approach. Front Genet. 2022 Jul

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: P43686 Human

SwissProt: P54775 Mouse

SwissProt: Q63570 Rat

Synonyms

26S protease regulatory subunit 6B antibody

26S proteasome AAA ATPase subunit RPT3 antibody

26S proteasome AAA-ATPase subunit RPT3 antibody

MB67 interacting protein antibody

MB67-interacting protein antibody

MIP224 antibody

Protease 26S subunit 6 antibody

Proteasome (prosome macropain) 26S subunit ATPase 4 antibody

Proteasome 19S S6 antibody

Proteasome 26S subunit ATPase 4 antibody

Expand

Images

  • Western blot analysis of Tbp7 on different lysates with Rabbit anti-Tbp7 antibody (<a href="/products/HA721435" style="font-weight: bold;text-decoration: underline;">HA721435</a>) at 1/1,000 dilution.<br /><br />Lane 1: K-562 cell lysate (20 µg/Lane)<br />Lane 2: HEK-293 cell lysate (20 µg/Lane)<br />Lane 3: HeLa cell lysate (20 µg/Lane)<br />Lane 4: U-2 OS cell lysate (20 µg/Lane)<br />Lane 5: HepG2 cell lysate (20 µg/Lane)<br />Lane 6: MCF7 cell lysate (20 µg/Lane)<br />Lane 7: Jurkat cell lysate (20 µg/Lane)<br />Lane 8: Mouse testis tissue lysate (40 µg/Lane)<br />Lane 9: Rat testis tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 47 kDa<br />Observed band size: 47 kDa<br /><br />Exposure time: 17 seconds;<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/HA721435" style="font-weight: bold;text-decoration: underline;">HA721435</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Tbp7 on different lysates with Rabbit anti-Tbp7 antibody (HA721435) at 1/1,000 dilution.

    Lane 1: K-562 cell lysate (20 µg/Lane)
    Lane 2: HEK-293 cell lysate (20 µg/Lane)
    Lane 3: HeLa cell lysate (20 µg/Lane)
    Lane 4: U-2 OS cell lysate (20 µg/Lane)
    Lane 5: HepG2 cell lysate (20 µg/Lane)
    Lane 6: MCF7 cell lysate (20 µg/Lane)
    Lane 7: Jurkat cell lysate (20 µg/Lane)
    Lane 8: Mouse testis tissue lysate (40 µg/Lane)
    Lane 9: Rat testis tissue lysate (40 µg/Lane)

    Predicted band size: 47 kDa
    Observed band size: 47 kDa

    Exposure time: 17 seconds;

    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 (HA721435) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Tbp7 antibody (<a href="/products/HA721435" style="font-weight: bold;text-decoration: underline;">HA721435</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/HA721435" style="font-weight: bold;text-decoration: underline;">HA721435</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 colon carcinoma tissue with Rabbit anti-Tbp7 antibody (HA721435) 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 (HA721435) 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 cerebellum tissue with Rabbit anti-Tbp7 antibody (<a href="/products/HA721435" style="font-weight: bold;text-decoration: underline;">HA721435</a>) at 1/2,000 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/HA721435" style="font-weight: bold;text-decoration: underline;">HA721435</a>) at 1/2,000 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 cerebellum tissue with Rabbit anti-Tbp7 antibody (HA721435) at 1/2,000 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 (HA721435) at 1/2,000 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 cerebellum tissue with Rabbit anti-Tbp7 antibody (<a href="/products/HA721435" style="font-weight: bold;text-decoration: underline;">HA721435</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/HA721435" style="font-weight: bold;text-decoration: underline;">HA721435</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 cerebellum tissue with Rabbit anti-Tbp7 antibody (HA721435) 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 (HA721435) 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 U-2 OS cells labeling Tbp7 with Rabbit anti-Tbp7 antibody (<a href="/products/HA721435" style="font-weight: bold;text-decoration: underline;">HA721435</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Tbp7 antibody (<a href="/products/HA721435" style="font-weight: bold;text-decoration: underline;">HA721435</a>) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of U-2 OS cells labeling Tbp7 with Rabbit anti-Tbp7 antibody (HA721435) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Tbp7 antibody (HA721435) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of U-2 OS cells labeling Tbp7.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721435" style="font-weight: bold;text-decoration: underline;">HA721435</a>, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of U-2 OS cells labeling Tbp7.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA721435, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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