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CD68 Recombinant Rabbit Monoclonal Antibody [PSH06-65] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751094

CD68 Recombinant Rabbit Monoclonal Antibody [PSH06-65] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • IHC-P

  • IF-Tissue

  • IHC-Fr

Reactivity

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

CD68 Recombinant Rabbit Monoclonal Antibody [PSH06-65] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within mouse CD68 aa 1-316.

Species Reactivity

Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, IF-Tissue, IHC-Fr

Molecular Weight

Predicted band size: 35 kDa

Positive Control

RAW264.7 cell lysate, M‑NFS‑60 cell lysate, J774A.1 cell lysate, Rat spleen tissue lysate, RAW264.7, mouse brain tissue, mouse intestine tissue, mouse spleen tissue, rat intestine tissue, rat spleen tissue.

Conjugation

unconjugated

Clone Number

PSH06-65

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:500

  • IF-Tissue

  • 1:500

  • IHC-Fr

  • 1:1,000

Target

Function

CD68 (Cluster of Differentiation 68) is a protein highly expressed by cells in the monocyte lineage (e.g., monocytic phagocytes, osteoclasts), by circulating macrophages, and by tissue macrophages (e.g., Kupffer cells, microglia). Other names or aliases for this gene in humans and other animals include: CD68 Molecule, CD68 Antigen, GP110, Macrosialin, Scavenger Receptor Class D, Member 1, SCARD1, and LAMP4. The mouse equivalent is known as "macrosialin".

Background References

1. Choudhary M et al. CD68: Potential Contributor to Inflammation and RPE Cell Dystrophy. Adv Exp Med Biol. 2023

2. Deng R et al. Periosteal CD68(+) F4/80(+) Macrophages Are Mechanosensitive for Cortical Bone Formation by Secretion and Activation of TGF-beta1. Adv Sci (Weinh). 2022 Jan

Subcellular Location

Endosome membrane, Lysosome membrane; Cell membrane.

UNIPROT

SwissProt: P31996 Mouse

Entrez Gene: 287435 Rat

Synonyms

CD 68 antibody

CD68 antibody

CD68 antigen antibody

CD68 molecule antibody

CD68_HUMAN antibody

DKFZp686M18236 antibody

gp11 antibody

Gp110 antibody

LAMP4 antibody

Macrophage antigen CD68 (microsialin) antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of CD68 on different lysates with Rabbit anti-CD68 antibody (<a href="/products/HA751094" style="font-weight: bold;text-decoration: underline;">HA751094</a>) at 1/5,000 dilution.<br /><br />Lane 1: RAW264.7 cell lysate (20 µg/Lane)<br />Lane 2: M‑NFS‑60 cell lysate (20 µg/Lane)<br />Lane 3: J774A.1 cell lysate (20 µg/Lane)<br />Lane 4: NIH/3T3 cell lysate (negative) (20 µg/Lane)<br />Lane 5: Neuro-2a cell lysate (negative) (20 µg/Lane)<br />Lane 6: Rat spleen tissue lysate (no heat) (40 µg/Lane)<br /><br />Notice: no heat means the lysate is not boiled.<br /><br />Predicted band size: 35 kDa<br />Observed band size: 90-200 kDa<br /><br />Exposure time: 12 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/HA751094" style="font-weight: bold;text-decoration: underline;">HA751094</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of CD68 on different lysates with Rabbit anti-CD68 antibody (HA751094) at 1/5,000 dilution.

    Lane 1: RAW264.7 cell lysate (20 µg/Lane)
    Lane 2: M‑NFS‑60 cell lysate (20 µg/Lane)
    Lane 3: J774A.1 cell lysate (20 µg/Lane)
    Lane 4: NIH/3T3 cell lysate (negative) (20 µg/Lane)
    Lane 5: Neuro-2a cell lysate (negative) (20 µg/Lane)
    Lane 6: Rat spleen tissue lysate (no heat) (40 µg/Lane)

    Notice: no heat means the lysate is not boiled.

    Predicted band size: 35 kDa
    Observed band size: 90-200 kDa

    Exposure time: 12 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 (HA751094) at 1/5,000 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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of CD68 on different lysates with Rabbit anti-CD68 antibody (<a href="/products/HA751094" style="font-weight: bold;text-decoration: underline;">HA751094</a>) at 1/5,000 dilution.<br /><br />Lane 1: RAW264.7 cell lysate<br />Lane 2: RAW264.7 cell lysate treated with deglycosylation<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 35 kDa<br />Observed band size: 60-100 kDa<br /><br />Exposure time: 10 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/HA751094" style="font-weight: bold;text-decoration: underline;">HA751094</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of CD68 on different lysates with Rabbit anti-CD68 antibody (HA751094) at 1/5,000 dilution.

    Lane 1: RAW264.7 cell lysate
    Lane 2: RAW264.7 cell lysate treated with deglycosylation

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 35 kDa
    Observed band size: 60-100 kDa

    Exposure time: 10 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 (HA751094) at 1/5,000 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.

  • Application: IHC-Fr<br /><br />Species: Mouse<br /><br />Site: Spleen<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1/1,000<br /><br />Antigen retrieval: Not required

    Application: IHC-Fr

    Species: Mouse

    Site: Spleen

    Sample: Frozen section

    Antibody concentration: 1/1,000

    Antigen retrieval: Not required

  • Application: IHC-Fr<br /><br />Species: Rat<br /><br />Site: Spleen<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1/1,000<br /><br />Antigen retrieval: Not required

    Application: IHC-Fr

    Species: Rat

    Site: Spleen

    Sample: Frozen section

    Antibody concentration: 1/1,000

    Antigen retrieval: Not required

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Immunocytochemistry analysis of RAW264.7 (positive) and Neuro-2a (negative) labeling CD68 with Rabbit anti-CD68 antibody (<a href="/products/HA751094" style="font-weight: bold;text-decoration: underline;">HA751094</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 Rabbit anti-CD68 antibody (<a href="/products/HA751094" style="font-weight: bold;text-decoration: underline;">HA751094</a>) at 1/100 dilution in 1% BSA in PBST 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. 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/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 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.

    ☑ Relative expression (RE)

    Immunocytochemistry analysis of RAW264.7 (positive) and Neuro-2a (negative) labeling CD68 with Rabbit anti-CD68 antibody (HA751094) 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 Rabbit anti-CD68 antibody (HA751094) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-CD68 antibody (<a href="/products/HA751094" style="font-weight: bold;text-decoration: underline;">HA751094</a>) at 1/500 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/HA751094" style="font-weight: bold;text-decoration: underline;">HA751094</a>) at 1/500 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 brain tissue with Rabbit anti-CD68 antibody (HA751094) at 1/500 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 (HA751094) at 1/500 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 intestine tissue with Rabbit anti-CD68 antibody (<a href="/products/HA751094" style="font-weight: bold;text-decoration: underline;">HA751094</a>) at 1/500 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/HA751094" style="font-weight: bold;text-decoration: underline;">HA751094</a>) at 1/500 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 intestine tissue with Rabbit anti-CD68 antibody (HA751094) at 1/500 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 (HA751094) at 1/500 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 spleen tissue with Rabbit anti-CD68 antibody (<a href="/products/HA751094" style="font-weight: bold;text-decoration: underline;">HA751094</a>) at 1/500 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/HA751094" style="font-weight: bold;text-decoration: underline;">HA751094</a>) at 1/500 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 spleen tissue with Rabbit anti-CD68 antibody (HA751094) at 1/500 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 (HA751094) at 1/500 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 intestine tissue with Rabbit anti-CD68 antibody (<a href="/products/HA751094" style="font-weight: bold;text-decoration: underline;">HA751094</a>) at 1/500 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/HA751094" style="font-weight: bold;text-decoration: underline;">HA751094</a>) at 1/500 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 intestine tissue with Rabbit anti-CD68 antibody (HA751094) at 1/500 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 (HA751094) at 1/500 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 spleen tissue with Rabbit anti-CD68 antibody (<a href="/products/HA751094" style="font-weight: bold;text-decoration: underline;">HA751094</a>) at 1/500 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/HA751094" style="font-weight: bold;text-decoration: underline;">HA751094</a>) at 1/500 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 spleen tissue with Rabbit anti-CD68 antibody (HA751094) at 1/500 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 (HA751094) at 1/500 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.

  • Application: IF-tissue<br /><br />Species: Mouse<br /><br />Site: Spleen<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/500

    Application: IF-tissue

    Species: Mouse

    Site: Spleen

    Sample: Paraffin-embedded section

    Antibody concentration: 1/500

  • Application: IF-tissue<br /><br />Species: Rat<br /><br />Site: Spleen<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/500

    Application: IF-tissue

    Species: Rat

    Site: Spleen

    Sample: Paraffin-embedded section

    Antibody concentration: 1/500

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