Overview

  • Product name
    Anti-GC1q R antibody [60.11]
    See all GC1q R primary antibodies
  • Description
    Mouse monoclonal [60.11] to GC1q R
  • Host species
    Mouse
  • Specificity
    This antibody recognizes the mature receptor at the amino terminus (aa 76-93), in the region of the receptor that binds C1q. It will block the C1q/gC1q-R interaction.
  • Tested applications
    Suitable for: Neutralising, IHC-P, WB, IP, ELISA, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Bacterial expressed recombinant full length protein GC1q R.

  • Positive control
    • This antibody gave a positive signal in Human tonsil tissue sections. This antibody gave a positive signal in the following Methanol fixed cell lines: HeLa.
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab24733 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Neutralising Use at an assay dependent concentration. PubMed: 9233640
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use a concentration of 5 µg/ml. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa).
IP 1/5000.
ELISA Use at an assay dependent concentration.
Flow Cyt Use 0.5µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use a concentration of 5 µg/ml.

Target

  • Function
    Binds to the globular "heads" of C1Q thus inhibiting C1 activation.
  • Sequence similarities
    Belongs to the MAM33 family.
  • Cellular localization
    Mitochondrion matrix. Nucleus. Might also be nuclear in some cell types.
  • Information by UniProt
  • Database links
  • Alternative names
    • ASF/SF2 associated protein p32 antibody
    • C1q globular domain binding protein antibody
    • C1qBP antibody
    • C1QBP_HUMAN antibody
    • Complement component 1 q subcomponent binding protein antibody
    • Complement component 1 Q subcomponent binding protein mitochondrial antibody
    • Complement component 1 Q subcomponent-binding protein, mitochondrial antibody
    • GC1Q R antibody
    • GC1q R protein antibody
    • GC1q-R protein antibody
    • GC1QBP antibody
    • GC1QR antibody
    • globular domain of, C1q, receptor for antibody
    • Glycoprotein gC1qBP antibody
    • HABP 1 antibody
    • HABP1 antibody
    • Hyaluronan binding protein 1 antibody
    • Hyaluronan-binding protein 1 antibody
    • Mitochondrial matrix protein p32 antibody
    • p32 antibody
    • p32 splicing factor antibody
    • p33 antibody
    • Pre mrna splicing factor SF2 P32 subunit precursor antibody
    • SF2p32 antibody
    • Splicing factor SF2 associated protein antibody
    see all

Images

  • All lanes : Anti-GC1q R antibody [60.11] (ab24733) at 5 µg/ml

    Lane 1 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate
    Lane 5 : Human liver tissue lysate - total protein (ab29889)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 33 kDa
    Observed band size: 30 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 68 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 3 minutes
  • Lane 1 : Mw standards
    Lanes 2-3 : Anti-GC1q R antibody [60.11] (ab24733) at 1/2000 dilution

    Lane 2 : 293T whole cell lysate at 20 µg
    Lane 3 : Hela whole cell lysate at 20 µg

    Secondary
    Lanes 2-3 : Alexa Fluor® 680 conjugated goat anti-mouse

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 33 kDa
    Observed band size: 32 kDa why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    The blot was blocked with 5% milk for 1 hour at 25°C prior to incubating with the primary antibody for 13 hours at 4°C.

    See Abreview

  • ab24733 used in Flow Cytometry.
    U937 cells were cultured in RPMI 10% FCS, treated with PMA (5nM) and, 6-12 hours later, cells were harvested, spinned and resuspended to 200,000 cells in 100µl. ab24733 used at 10µg/ml for 30 minutes at 4°C. A phycoerythrin conjugated goat anti-mouse polyclonal was used as the secondary antibody at a 1/100 dilution.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab24733 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab24733, 0.5μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150117) at 1/4000 dilution for 30 min at 22°C.

    Isotype control antibody (black line) was mouse IgG1 [15-6E10A7] (ab170190, 0.5μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

    This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

  • IHC image of ab24733 staining in human tonsil formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab24733, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • ICC/IF image of ab24733 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab24733 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:
  • Gao H  et al. Elevated HABP1 protein expression correlates with progression and poor survival in patients with gastric cancer. Onco Targets Ther 9:6711-6718 (2016). WB ; Human . Read more (PubMed: 27826197) »
  • Ansseau E  et al. Homologous Transcription Factors DUX4 and DUX4c Associate with Cytoplasmic Proteins during Muscle Differentiation. PLoS One 11:e0146893 (2016). IP . Read more (PubMed: 26816005) »
See all 6 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (gradient 4% to 12%)
Sample
Pig Cell lysate - whole cell (PK15)
Specification
PK15
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 23°C

Mr. Guy Kouokam

Verified customer

Submitted Jan 23 2015

Answer

Thank you for your reply.

I will recommend you to use the next antibody as an isotype control for Functional Assays:

https://www.abcam.com/Mouse-IgG1-kappa-monoclonal-MG1-45-Isotype-control-ab18447.html

As it is suitable for these kinds of assays it is therefore free of sodium azide.

I hope this helps. If not please let me know and I will be happy to assist you further.

Read More

Answer

Vielen Dank für Ihre Antwort und für diese weiteren Informationen.

Es tut mir leid zu hören, dass unsere Vorschläge in diesem Fall nicht zu einer Verbesserung Ihrer Ergebnisse geführt haben. Ich weiß, dass Sie viel Zeit für diese Experimente verwendet haben, und stelle Ihnen daher gerne den ab101267 Anti-GC1q R antibody als kostenlosen Ersatz aus (vorausgesetzt, das Produkt wurde in den letzten 180 Tagen gekauft). Dafür benötige ich jedoch ihre ursprüngliche Bestellnummer oder das Datum der Bestellung.


Vielen Dank für Ihre Kooperation und Hilfsbereitschaft. Ich freue mich bald wieder von Ihnen zu hören.

Read More

Question

1) Abcam product code ab24733
2) Abcam order reference number or product batch number
3) Description of the problem: no IP
4) Sample preparation:
Type of sample (whole cell lysates, fraction, recombinant protein…): whole Hela cell lysate
Lysis buffer Tris pH8,0; NP40; NaCl
Protease inhibitors: Roche
Phosphatase inhibitors Roche
Amount of Lysate IPed ug or number of cells 4 000 000 cells per IP
Lysate precleared with matrix : no
Positive control: yes
Negative control: yes
5) IP antibody: ab24733
Amount/concentration or dilution used 1:5000
Conjugation:
Antibody noncovalently bound to matrix before incubation with lysate sample?
Antibody-lysate incubation time: over night
Incubation temperature 4°C
6) IP Matrix (e. beads): beads
Type of matrix Dynabeads Protein G and A (Invitrogen)
Amount of matrix 1,5 mg
7) Washing after antibody-lysate incubation:
Buffer Lysisbuffer
Number of washes 5 times
8) IP analysis:
Verification method: e.g reprobe in western blotting: WB
Describe verification method briefly: Input, IP, non-bound fraction
OR if Western blotting reprobe performed please indicate the following:
Reducing agent: Lämmlibuffer (SDS, beta 2-mercaptoethanol)
Boiling for ≥5 min? 5 min
Protein loaded ug/lane or cells/lane 2 000 000 cells per IP, 200 000 cells Input and non-bound fraction
Percentage of gel: 10,5%
Type of membrane: Hybond ECL (GE)
Protein transfer verified yes
Blocking agent and concentration 5% Milk in TBST
Blocking time 1h
Blocking temperature RT
Primary antibody (If more than one was used, describe in “additional notes”) ab24733
Concentration or dilution 1:200
Diluent buffer 5% BSA in TBST
Incubation time over night
Incubation temperature: RT
Secondary antibody: Anti-Mouse IgG-HRP
Species: Mouse
Reacts against: ab24733
Concentration or dilution 1:5000
Diluent buffer 5% Milk in TBST
Incubation time 1h
Incubation temperature: RT
Fluorochrome or enzyme conjugate: HRP
Washing after primary and secondary antibodies:
Buffer: TBST
Number of washes 3
Detection method ECL
Was the method successfully used to detect protein from non-IPed samples? yes
Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.

Read More
Answer

Vielen Dank für Ihre Anfrage und dafür, dass Sie sich die Zeit genommen haben, unseren Fragebogen auszufüllen.

Es tut mir leid, dass Sie Probleme mit unserem Antikörper hatten.

Um Ihr Problem zu lösen, möchte ich die folgenden Vorschläge zu Veränderung Ihres Protokolls machen:

1. Mir ist aufgefallen, dass die empfohlene Starkonzentration für all unsere anderen Applikationen 5ug/ml beträgt, und dies ist doch ausgesprochen ungewöhnlich, da IP normalerweise eine höhere Konzentration benötigt als z.B. ein WB. Ich vermute daher, dass es sich hier um einen Datenblattfehler handelt, und habe unser Labor zur Rückmeldung kontaktiert. Ich möchte Ihnen daher empfehlen, die Konzentration ebenfalls auf 5 ug/ml zu erhöhen.

2.Um den Antikörpern die Bindung an die Beads zu erleichtern, möchte ich Vorschlagen, erst die Antikörper an die Beads zu koppeln ( Inkubation für 2h), und dann erst mit dem Lysat zu behandeln.



Bitte lassen Sie mich wissen, ob ich Ihnen helfen konnte. Wir bieten unseren Kunden gerne einen kostenlosen Ersatz, eine Gutschrift oder eine Erstattung an, wenn das Produkt wie auf dem Datenblatt beschrieben behandelt und innerhalb von 180 Tagen nach Erhalt reklamiert wurde.

Zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (Human Cell (U-937 (ATCC: CRL-1593.2) histiocytic l)
Specification
Human Cell (U-937 (ATCC: CRL-1593.2) histiocytic l
Preparation
Cell harvesting/tissue preparation method: U937 cells were culture in RPMI 10% FCS, treated with PMA (5nM) and, 6-12 hours later, cells were harvested, spinned and resuspend to 200 000cells in 100ul
Sample buffer: PBS + 0.1% FCS
Fixation
No fixation
Permeabilization
No
Gating Strategy
FS - SS

Mr. David Sanchez Martin

Verified customer

Submitted Jan 24 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Hela and 293T)
Loading amount
20 µg
Specification
Hela and 293T
Gel Running Conditions
Reduced Denaturing (4-12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted May 23 2008

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

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